Chambers_et_al_-_Flavivirus_Genome_Organization_Expession_and_Replication_-_AnnuRevMicrobiol1990.pdf

8/10/2019 Chambers_et_al_-_Flavivirus_Genome_Organization_Expession_and_Replication_-_AnnuRevMicrobiol1990.pdf

1/40

Rev Microbo. 990 9Cpyght 990 l Reews ll hs eseed

FLA lIVIRUS GENOME

ORGJ\NIZATION, XPRESSION

AND RELICAION

haJ

habr, hangS

Hahn, Rard ar, andhar M R

Depatment of Molecua Microbioogy, Wahington Univrit Schoo of Mdicin,

ox 8230, 60 South uclid Avenue St ouis Misouri 3110-1093

Y WORDS poite-ta RNA i. potei poesig aima gyopoteipoteae

CONTENTS

ORW................................................................. 650

INTRODUCION TO AVVRUS ................................. 650assa a Nau 650pa as Disas As 651pai i uud s 65

ORGANIZAION AND RANSAION OF HE ENOME RNA ................ 65Ga aus ............................ ............................ 65 a s a i S a 65

PROOYC PROCSSIN O H IRA POLYPROIN.............. 655 SuUa[ Ps 65T ouua Po . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

PROPERE O VRA PROENS ......................................... 660Suua Po . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 660Nnual Pn................................................................. 66

VRION SUUE AND ASSML.................................... 61

ONSERVE RNA SEQUENES AN SRUURES................... 6 an 3' nal Snay Se ................................ 67s a pa Squs 675

RN REIAION ............... .................................... 675

Ie ae: Fuaao Owalo C Deptmeto e Boqumia e Bioogaoa, Rio e Jio, CEP 21040, Baz

9

0047/0/0004$0.00

Annu.Rev.Microbiol.1990.44:649688

.Downloadedfromwww.annualreviews.org

by

UniversityofMichigan-AnnA

rboron02/26/11.Forpersonaluseonly.


2/40

50 CHAMB E A

OMPARAIV ASPTS.... .. 677ene gene ng vve . .... .. ............ ...... 678iiitis th sitit iss . 679

CNCLUDNG MAK AND FUU RPCS................... 679

OVERVIEW

vvuses re sm enveoped nm vruses contnng snge postvestrnd genomc N Mode studes on tese vruses begn wt te dscovey ney centuy go tt te dsese yeow feve ws cused by fltebe gent nd trnsmtted to umns by mosutoes (see 0) Tsevew summes ecent deveopments n vvus moecur boogy

empsng te v genome stcture nd te epresson nd possbefunctons of te vr protens Hggts of te st sever yers ncude (a) compete mp of te flvvus-encoe potens nd n emegng pctue ofte pocessng events nvoved n ter poucton etensve seuencecomprsons owng te dentfcton of potenty mportnt stuctumotfs n te enomc Ns n encoe ptens te evton of stuctu mode for te mor von enveope proten ( te ecovey ofnfectous vvus RN fom moecury coned DN In ctng tetetue, peference s been gven to recenty pubsed mte. Te

ee s efee to two boos 13 fo compeensve revews ofele wo

NTDUCTIN TO FLARUS BLOGY

Classficaton and Lfe Cycle Nature

Te Flaviviridae (fom te tnavus. or "yeow referng to te protote us eo feer us s reent estbse s serte fm78) stnct from te Togaviridae nd current ncudes 68 memers 1 .Te mjort re rtropodboe trnsmtted to vertertes croncnfecte mosquto or tc vectors However sotes from bts n rodentstout nown nsect vectors ve so been entfed Te fe cce of teopo-boe flvvruses nvoes compe retonsps mong nsectvectors verebrte reservors umns n te envronment (revewed n 19.

vvses demonstrte ntgenc rossrectvt tt s strongest n teemggutnton nbton ss H te gest secfct s sown nneuzton (NT sss 18. Crossneutrton usng pocon ntser s te bss for te serton of vvruses nto eg ntgenc compees members of n serocome common sre rnge of oogcopees (gure e four mosquto-boe comees ncue teJnese encets (J for vvrus bevtons see gure groupNt Ugnd S, nd te DEN groups . Te tc-boe comees ncude teTBE n te Tyuen groups. Compees wtout dentfed tropod vec-



by




3/40

AVIVIUS GNOM STUCTU & XPSSION 61

Angenic mplx p b or s wh thopd valab u (r of b vt

Pe cic N-mal Ca

Tck-eit(12) m

1 7 .1

HEE 127,27a,88o B 6) o Bo 0 M 10IS7 1IIS12IS760

KjK M 247,49 48 ry y i phs E M 4131 101WstNeWN 15.16121172 2 0,111,17.171.17273

Tliy ( yuly

Ny5 ay '

U gnd M"4 (N) 2,15

Dgetyp 2DEN2 4S'II84 373818077 187 (N) 4"t4DN4 M 71

M 5)

U "

17 Ylow fvr YF) ' 421131 335758132133

r 1 avivirs antigenc complexes, arthropod vecors and sequence data Anigencgoups ae tken fom Refeence 1 Atopod vectors: tc; mosquo unnownastes pod vetos fo some members o tese oups ave no been dentfed. eunroued vivirses ncude mosuito- and tictransmed vises as wel as some w nonown veco Incuded ae teratue citaons of pused avvs poen o nucec acdsequence da

ors includ ssocd Ro Brvo group nd odnssocdModoc goup. In ddion 7 vuss icludng F dmons nurli

on propis do no llow clssicion win s srocomplxs. Ronsps mong vivus sd upon comprson o nuclccid nd proin sunc d, closly ollow os dducd rom crossneutralzation tests sing polyclonal atsera (see below). However, elaonsips do no i rdonl clssicion cn dnd pop lvl usng monoclonl niods nd lv sologc cnius 8

Iran D Agn

log umns r primrly dnd oss o mnnnc o lvivuss n r nurl cycl 8) rropod-o vviruss r sponslor signicn umn nd nml dis worldwd 10 1 8) Clincl dss is xpssd s v ncplis o morrgic vr ndncluds svrl niis o mjor glol conc ncluding yllow vr(11) dngu v w s ssocd dngu morrgc fv (D ndsoc syndrom (D (rvwd n nd pns ncpls BE Kynur os dss WN E nd ME o imporngns o gonl ndmic o pdmc dss 10) Dsurncs in vcovrr quilirum v rsuld in sgnicn ngonl spdo s vss Worldwd dico o vivirs pogns s unllycus y r mnnd n nml rsors nd cn nsovrlynsmid in ropods. us r vccnion s only vll o F



by




4/40

65 CHAMBS T A

uin he aenuae D rain (6 an for BE an JE uin inaivaeviru (reviewe in 6

Rpcan n Cutud Cell erive fro any aalian avian, an arhropo oure anuppor faviviu repliaion in ell ulure (0 he early even in faviviru infeion are no well haraerize an ellular reepor for aviviruehave no been ienifie Eviene ue ha he viru ener ell byreepor-eiae enoyoi (, preuably eiae by he viion envelope roein (E an plaa ebrane reepor n aiion in hepreene of ubneuralizin oneraion of aniboy reepor alo

eiae aahen an upake ( he laer eny ehani, erey hm ay play a role in evelopen of DHD in equenial infeion wih ifferen DE eroype (reviewe in 66 Ulrauural uie of W infeion of PD l ell an of anU infeion of Vero ell have loalize inle virion an virion areae o lahrin-oae pi on he ell urfae upake ino oae veilerapily follow aahen ( , Virion are laer foun in unoaeprelyooal veile where an aiaalyze ebrane fuion i houho releae he nuleoapi ino he yopla (5 Conien wih hi

releae a onforaional hane in he viral E proein (ee below poiblyipoan for fuion our a low p (55 Ai pH an alo pooefuion of W virion wih lipooal ebrane or a he plaa ebrane alhouh hi oe of eny oe no lea o prouive infeion (5 Ularuural analyi of he enry of oquiopaae E an DEvirue ino oquio ell or huan onoye ue ha ire plaaebrane peneraion an our (6 alhouh wheher hi iniiae a pouiv infion i no l

he unoain of virion, ranlaion of he infein virion RA, an he

iniiaion of RA repliaion have no been uie Viral RA ynhei anbe eee a hree o ix hour wih he releae of infeiou viru beinnina appoxiaely hour Alhouh rowh kinei appear iilar in verebrae an ahropo ell axial ier in ifferen ell ype an vyonierably ( ven urin he peak of viu prouion a ajorinhibiion of ho arooleular ynhei i no obeve ( 6 Inverebrae ell raai yopahi an ulauural hane an beoberve inluin vauolaion an proliferaion of inaellular ebrane1) infeion i oonly yoial alhouh oe verebrae el yeo no how hee effe an beoe hronialy infee Arhopo ellay eonrae yopahi effe inuin ynyiu foaion (reviewe in 6 enerally infeion are nonyoahi ( an erie



by




5/40

AVIVUS GNOM STUCTU XPSSON 653

infections e more easily established in athopod cells than in vetebratecells.

RGANZATN AND TRANATN F THGNM RNA

Gnrl Fturs

The genome RNA of avivirses is single-sanded and approximately 11kilobases i legth (reviewed i 9 134, 76, 77) The geomic RNA isifectious d thus of positive polrity ecodig the vrl proteis ecessryfor RNA replicatio. Geoe-legth RNAs apper to be he oly virus

specic NA molecles in avivirs-nfected cells. The genomic RNA hasa type cap at its 5' end (mGpppAmp) and lacks cap-associated and intealbasemethlated adeine residues. The 5 cap is followed by the coserveddiucleotide sequece AG Geoic RNAs of osquito-boe avivirusesapper to ack a 3'-temial poy(A) tract 9 34, 7, 77) and insteadteriate ith the coserved diucleotide CU. TBE RNA probably terminates wih the dinucleotide AC (100, 127a), bt the presence of additioalseqences or a poly(A) tact have not bee rigoously exclded C. Mndl &F X Heiz, personal commnicatio).

The ot otable featre of the avivirs geoe is the presece of aope readig frae (ORF) of over 0,000 bases. ORFs for dieret avivirses (startig at the rst Met residue rage from 0,158 bases (DEN4) to0,302 bases (MVE) and ecode polypoteins of 3,386 to 3,434 amio acids

(compiled in Figre 2). No other coseed ORFs have been identied ineiher the geomic sese RNA or its coplement. Flaig the long ORF re5 (from 9 to 3 bases i legh) ad 3 ocodig regions (Figre ) The3 ocodg regio of TBE (114 bases) appers to be draatically shorerha that f ay of the mosqito-boe viuses (from 385 to 585 bases in

length) an lacks a nmber of RNA seqences ad stctres common to hemosquito-boe viuses (see below).

Aother iteresting featre of avivis geome RNAs is their high prinecotent and low CG ad UA doblet eqecies, the sigificace of which isucler. As discussed i detail elsewhere (133, 134), these orado diucleotide freqecies ad the resultig biased codo usage i he ORF ayinuence translation of the lavivirs genome ad reect the adaptatio ofthese vires to growth their host evioments.

Tnsln SrgThe accuulated potei ad ucleic acid sequence data for many aviviruses(compiled in Figre and discussed below) have shaped our crrent view of



by




6/40

6 CHAMBS T A

Favivirs (stn) 5' NC

DEl >67JAM 96DPR -963 ?94

DE4 01 (OS98) 95 Beijin- 95KUN >7 96SLE 9TE(W) 1W 96

YF17 11YFAsibi 1

OR

10,1701

10110,9610,710,990,0

1041090

1010

3' NCgnome

Souelen

1044 107 7,8 0,2 60

14

0644 9790 10976 17 1096 66

>90 >10664 8> >1096 ,90

166114 10,4 99100574 10,960 1,16,17

1,1,11 10,6 111 0 62 57

gu F1 gom z L g o 5' oo go (5 N, og op ig m (O) ooig go NC; u opcoo miig O o

viral proei yhei he orer of proei eoe i he o OR i5 Cpr(E NS2ANS2NS3NSANSNS3 he ruural proei C (api; he C preuror i ale ahore C) (ebraehe preuror i alle pr a (evelope are eoe i he quarerof he eoe a he ee for he o ruural proei ae loae i hereaier he o ruural proei ilue he lare hihly oereproei 1 a a he four al hyrophobi poei AB A a B For a erpio of he proei oeaure uehere a i reaiohip o previo e eiaio ee he followireferee (1 1 1

-erial equee aa for he E YF W a E C proei howha ralaio a bei wih he fr AUG i he o ORF Oy of he1 1 avivirue for whih aa are avalable have a purie i he poiiorelaive o hi AUG whih i preferre for eukaryoi ralaio iiiaioCC(GCCAUGG (8 86 he oquio-boe virue have a eoifrae AUG bewee 1 a 1 oo owrea fro he fir AUGwih a A reiue a he 3 poiio or W eria equee aalyiof virio-aoiae C proei (ee beow (16 ue ha hi AUG ayalo erve a a ralaio iiiaio ie I BE a aiioal hor OR of oo i pree ' o he lo ORF C al & X Heiz peroalouiaio 1 188 ralaio eriaio of he lo ORF ha oro eriaio oo preferee all hree are repreee UAA (DEU E W E UAG (DE a UGA (

everal experieal approahe le o he ueio ha oe faviviruproei ih be proue fro he viral mRA by iea iiiaio of

alaio Aeaive iereaio of hee experie have bee iue elewhere ( 1 1 he ue view ive he preee of a ieORF eoi a ea 10 avivirpeifi proei i ha ralaioiiiae ear he 5' e of he eoe a ha iiviual vial proei are



by




7/40

AVIVIRUS GENOME STRUCTURE & EXPRESSION 6

podued by poeoy T ew uppoed by e aaabe poeequee daa ue ad , eee aao o eom RNA eebeow, e detifcato of i moeua we poypoei i ifectede 2 , 26, ad e ee aao mapp epeme u KUNiected o ce, wic demoaed a e ia poei ae yteiedi te od pediced by te eome equece .

PT YT PSS T ALPOL YPROEIN

aed o N ad C-emia poe eqece daa ad fom e aime ofomooou equece (iue , poeoyic ceaae epaai te ia

poypepide a be caified io diic oup. Te N emii of p, Ead N foow peded aae eaae e 68 obued by eema yopo eo o aoe , p, a epeey Aa equece ao peede e N emiu of N uei a iydopob poe poceed aoao w edopam euumR membae Te N emiu of N2A foow a eaae e defied bye equece a-Aa =e, , , A, Ap at fufi e e o a aae e owee e e a a eue upeamydopob eo 8 . A poeae o oe pey a ee oed

fo ti caae 0 aou ceaae by iaae a ot beeexcuded Addiioa ceaae wii e N2A eio a bee ifeed 2,02, 0 ee beow, bu e ceaae ie ae o bee caacteied.Ceaae e eea te N emi o N2, N NA, ad N aey oeed amo ae Fue Te ou ae wo baamo ad edue LyA, AA, o Ay o oaoay aeA N ad N N ad ae uuay aed by o idecaiamio acid, mo commoy y, e, o Aa iue . A aiieoded peae wa ypoezed o edae uc eaae 33 a a

ey ommo o eea oe RNA e eewed 87, ad ee daaubaa bee ae peeed beow.

Aou oou poo o aaabe o may o e moeua eei ai poypoe poe eem ey a aao aoatio wi e ou R RR, aocato, ceaae, acquiiio of ecoec poei topooy, aemby of e epicaio compexe, ad iiomopoe ae a oey eoeed pomea Te oowmode ue aemp o put uey aaabe daa io a efamewo. ei w aao of e C poe, iaae ceaae

medae e pmay poe ee o ua poe peuoTe ae poe oa a -emia ydopob doma a a aa itea ia equece fo aoao of pM ito e R ume, wee



by




8/40

66 HAMRS A

S-ik cleavages (R lumn)

CM ME ENS! NS4A-NS4BDI LPTFHTTR TPRCVG GVOGCVD JFHTT =Jl J V.NEM AJFH JMVV JOMVD TJF VYMVV FVQJOM JNM

AYJMNF AY ANVHJOA VJNYM VJVLN J J FAF VAY.FNM ANVHOA VGLV!NGL L. AAY AVQ.W = ! VA VA1 VVI

VTLLMTAT YATH VA.OVAV AGVNL

leavages afer dibasc des (cyoplasc)

vv h Cv C l-2 2-3 3- 5

I ---JM N-----JAM JN QJ N J N-----JLCLMD N--J KGSRRSWPLNE QVQJWD G GGG

J QNN NNJTE JW F.A K BJ J J . QJ NNJ J . INSR.GGTRS HPNGJSWPASE GHSW AF ONJWE Y.VWD FAV .Y ----HOV FJ A. FJ T.

TE Q-ADM HJ OV YSJL

Or cleavag

MNt MM NS-NSA dibic, delay nov slgtl d

pdas. yol Iw Iw

I MJNNQ DJV EENAJEVO MJNQR JV D MJNNQ JV MJNQ J NQAJ

J M RRJ LQOAJFNEMV MJ J G SS!SITQT SLSQA!OI

J VQH EVTAJVAM

J TVQNAJYNOMY MJ J HLVAJTE MJ= SRSIPS QGSMV!ONG

re 3 Fliis clee sites_ Seqences sonin polpotein clee sites arrowsfo seel liises e line_ Alinments bse on N-teminl n C-teminl poteinseqence t (ndelnn fom nmbe of iffeent iises (Fie 1) Stin-scificieences sonin the clee sites wee note bt not shown hee The most skinsin-specic iffeence ws the seqence RGG t the to postions pecen the NSI-Aclee site in the Beijin-I stin of E ()_ n compilin these t pefeence ws ien tothe st l-lenth pbishe seqence fo ech eot_ Refeences e s follows DENI (10)

DEN (0); DEN (1); DEN (97, 190) JE (17); KUN () E ( 90) SLE (1); (11 1117) (1) E (99, 1)_



by




9/40

AVIVIRUS GENOME STRUCTURE & EXPRESSION 67

Pocessng o he spoproei

NC

ORF 3'NCS' _ cap

sRuURAy

NONSTRUCTURAL

+ + + ** -

o0 :; * LI f 1 1nc M E

.'

E on

ssem?

vnreese?

NSI-2A NS2B

**

NS NSA

NS3 NS4A4B NS5

+ ?

mNS4A NS4B

signala, rapd lumen

dbasic aid ytlasm dbsc dlad? ctoplm

U dibac deled lumenV novel, lightly deled lmen

r Schematc of te poc vt or aviv polypoei T op dpct tev genome the stct n nonsctl ein coing egions he c ttie 'secon stcte n the 5' ue ego e ce Boxe bow hegenome nice ecsos n mte po ad the pooytc ocessng cscedd box) stcl pteis; op box) nonstct po Blk b eesentogo tets o aged amo aid rk idae pobale N-ked gyoyato te fo F S te ext fo a diuo o te daa ppo th mol

the -pr ceavage and core glycosylation of pr occur Secondary cleavage of ao o a pM ay o o ay ada a a dud mo a ow. At the termnus of pr,adjacent hydrophobc setches nterrupted by a charged resdue act as atop-af qu fo pM ad a a ga qu fo aoao of po. Two ada ydopo qu at t mu o the

act simlarly fo po op-af ad S aoao Taoaoof S to t um of E foowd y aag of -S a oglycosylatn o S he S2A2B cleavage s mediated the cytosol by aviral proteae. leavage o S appears to be delayed or some lavivru ad ypozd to ou lumen of a ua omatmt.

Smaller S-Arelated forms have been detected suggesting poaa aag w the S go pror to the cleavage generatingt tmu of S . og of S2B-345 go popo to bemedated pmarily by a vral prtease; the S2A-2B S2B-, S4A, and

SB ee ou yoo d gaa ga the SB Nu. T cleavage strategy s consstent wt the cytoplasmc location ofS and S w a pumd yma components of he vral



by




10/40

68 AMR T A

repiase and it suggests tat te ydropoi NSA NSB NS4A andNS4B roteins ma be membraneassoiated, eras via membranesnnin domins Te exermenta dt sortn tis mode re smm

red eowhe rucura Proen

Anaysis o te poessing o te sttura region enoding C pr and invivo and in vitro indiates a reqirement or membranes and intat sinaseqenes. In vitro tanston o TB vrion NA as sown tat rodtiono vira C and proteins in retioye sates reqires a rtiate ratonrom Kres asites es and tat te ativity is destroyed by detergenttreatment 8 9) Tese oservations ave een onrmed or DN4

0) WN ) and YF ) y transation o transripts ontainin tetutura ene seqenes in retiuoyte ysates wit anine panreati miosomes Ctermin anysis o te WN Creted rotein rded in teresene o memranes ad anored C) revas an 8amino aidydropoi sement at te C temnus not present in iion C potein ).Te orresponding reion in 4 is neessary or oret expression o rin vitr deetion o 0 amino aids rom tis seqene or interuption byinseion o a nononseatie aio aid preents yosyation transoation nd orret eavae o r 0) Simiry deetion o te

omooous YF sequene ters te rreted n vitro rdts 37).Ceavae o te W pr and reated proteins durin eree tansation inte presene o membranes ors at te same signaase sites used in inetedes ) Furtemore te ydropobi sequene preedin te YF protein is neessay or ansoation o is protein in ito )

esuts onsistent wit signaasemediated roessing o te stuturarotein reion ve so been obined in avivrusineted es and intransin exprssion sysems sin vainia vs reominants N4strutra proteins an e expressed in te asene o downstream nonstrt

ra seqenes 8 4). eombinants ain te signa seqene preedinte protein do not exress ts rotein atoug sstitution wit aeteroogous sina sequene aows expression ). Frermore iniitiono KUN strtr reion proessn in e tre is oserved in te reseneo te ene ano treodrxyene wi is known to nibtsinaase eaae wen inoporated into sina peptides ).

he onrucura Proen

OSS O S O Anayses o NS proessing in avivirusineted es and in transien expresson systems indiat tat a sinasequene preedin te N temins s required and tat NSA may pay



by




11/40

A VIVIRU GNOM TRUTUR & XPRION 659

a roe in NS poessing n es inee wih DN4vainia eombinanse signa sequene bu no e puaive sop-anse sequene in e Cerminus o e poein is equie o NS epession 8 4) n YF

inee es eviene o an NS2A peuso o NS (2) sugess ha eNS2A-B v or iv vs in NS2A rion our ror oe NS-2A eavage NS2A as no been ieniie o DN2 8 oB (9) bu in inee es an aiiona 56-iaon () gyosyae NS Aeae poein (NS ) is pesen an seee wih simiaineis o hose o h 48 NS poein (02) A roe or NS2A in eNS 2A v is so sust by suis wi DN4vini rominns n i progressiv C-ermin eions o NS2A reion enrae uneave bu gyosyae NS -2Aeae poeins (4) NS gyosya

ion may aso be impoan o NS 2A eavage beause uniamyin eamen aes he pouion an seeion o NS eae oeins (02) angeneaes aeran evage pous o he YF NS -A poypoeins ()

PROCESSING OF THE NS2B-345 REION Comparaive sequene aa ( 46) (see seion on popeies o via eins beow) an een in vio anin vivo suies impiae he N-emina hi o NS as a seine poeaseesponsibe o meiain some o e nonsuuaegion eavages haou e ibas resiues (see seion on NS3 propeies) ansaion o

NA ansips in eiuoye ysaes sowe ha poypoeins onaininge popose NS poeae omain ae apabe o iuininseniive evage a e aueni NS2A2B an NS2B eavage sies an a amino aisubsiuion o he propose aivesie esiues in e NS proease omainen abis pessing (T Chames C ei A aoui D Cour F Baan J Feei & C ie submie) Ceavagea he N ermini o NS4A an NS5 wh ous e simia oube basiava sies s no y been onvininy emonsrae in viro

Daa om suies o via poeins synhesie in inee es ae aso

onsisen wih his pessing sheme an impiae a seine poease osome o he nonsuua poein eavages DN2speii peins o appan moeur weih () 0020 whih aumuae in he pesene oN-4osy-penyaany-oomeyeone an ZnC apea o b prusos o maer proeins (26) T arginin anaog anavanin ars eepession DN2 NS an poeins o 5 an 0 ae eee(25) Sevea YF-speii poypeins an be eee using egion-speiipoyona anisea (21). ese have he preie s o NS 4- (0) NS4AB-5 (40 ) NS4B5 (0 ) NS -4A (0 ) an NS4AB

( 9 ) ese aa sugges ha siespeii eavages ou on he nonsuu pypoein a e popose oube basi sies

he N-eina mino ai sequenes o KUN an YF NS4B suges



by




12/40

0 CHMBE E

snse m te ts eve 20 47) tou membne nvolvement s not been nvestted eementy. Ts ntrn obsevtonsests tt te leve event enetn te N temns o N os n

te men o te nd tt NA nd N ontn membne-snnnomns.n smmy oessn o te lavvs olten nvolves emkbe

oesson o eves medted by oteses o bot ost nd v on.Ate tnston o te noed C doob squene te vvsosome esumb beomes ssoted wt te nd tnston o v otens s ke to be membessoted. ute eements to mny o te dets o oessn nd to eludte te tu membnetoooes o vvus otens e ety needed.

PPS F PTS

Poessn o te vvrus ooten enetes t est 0 membnessoted v rotens (tee sttu seven nonstut otens) ndwo polyppi laag fagmn. Th followng on mmariz rent nomton edn te stutue mtton nd ossbe ntons otese oyetdes (Fe ).

tuctual PotenT CASD ROTN Te C roten esent n vons s sm (edtedW 214 kd) y ostvey ed oten 27% Ls+A o YF) 1 34) w oms stutu omonent o te nueosd Fvvus Coten sequene omooy s ow bt eons o ydoob nd ydo mno ds e onserved ndn C-temn ydoob domntt s mmedtey eeded by do eon nd enthyropho ron (99 h N-tn on ontns ydo seton tt n mosquto-boe vvuses s nteuted by doob nsetnot esent n TB

The vvus C oten ests numbe o deent oms de to botN- nd C-temn modtons Ntemn sequene dt (Fue 3) ndte tt te nttn metonne s emoved esumb b eu mnoetdse te ossbe Netyted N termn wod not ve beendeteted n tese stdes n t C rotens etnn te nttn metonneve been deteted o WN ) nd L () nd some WN C otens soben t te seond nme metonne t oston 1 o te F 1 1 2 1 ) .Fo vrion C otens o W N nd UN te C temnus s been ostonede dobe bs eve ste 18 mno d esdues om te Ntermnus (Fue 3) 121 48)

Deenes n te eetrooet mton o nte C otens



by




13/40

rne ofN c

%fav pced hydbi

P

a ril suesMW ava

YF)

C 12.3-137 M

vC 10.5117 M 7

rM 18.1-190 SI

S 39

pr 98107 SI

D 1

M 82-85 L S 8

533-3 S S 41

SI 390 S ?

23725. ? 56

1381

S3 68 I

39

1616 S 56

2659 S 49

NS 103-1 39

! -

t membr-hrge sa(F) d?

+25 y

26

+5 y

1

y

5 y

7 ?

+ ?

7 ?

0

2 ?

3 ?

1

N

i

? d e

n n; ba N U kn

_

I tal

y I . =

yI 6

dl

i - ! i ! c 12_ y mjI

mn s

(


14/40

662 AMR T A

von-assoated C otens and C otens odued n e-ee sstemsave een oseved. ntaeua KU C oten mates aste and as att etde omaed wt von asd (6). In ontast o W and

TB naeua C oten mates moe sowy tan te von-assoatedom and aks a tt etde (9 ). eent te deene etweenntaeua and von-assoated W C oten was not deteted ut teom odued n vto n te esene o memanes w ontans teC-temna doo doman, mated sowe tan te vonassoatedom akn ts doman (2). Sma esuts wee oseved o te TB Coten odued y e-ee tansaton atou sequene data was nototaned (58). Te C oten as een dut to detet n DN4 and YFeee tansaton studes ( 3. Te ntaeua YF oten de

teted ate sot useaen aso mates sowe tan te vonassoated om atou te stutua deene as not een aateed(2) A TB vaant Toan san ZZ/9) ontans a von-assoated Cten tat mates aste tan tse ote TB stans oss eauseo a nononsevatve amno ad susttuton (56 Te ese oes o and Ctemna modatons n tese eetooet deenes and n teunton o te C oten eman nea

PRO Te M oten s a ooten euso edted

MW 8.9. kd moded aodate addton) to te stutuaoten M . Ts euso undeoes a deaed eavae to om M and teN-tena segment, te ate o s unnown. Ts eavage sesumay ned to vus matuaton o eease as M and M e ound onntaeua and extaeua vons esetvey see eow)

temna amno ad sequene data Fue 3 ndate tat te temnus o M mmedate oows a snaase ste ( 2) ontuted te temnus o te asd oten. Te N tenus o te otenmmedatey oows a a o as amno ads eeved to eesent aeavae ste o ete a va o ost otease. Avaae C-temna sequenedata suest no ute tunaton o ts oten Fue 3).

e Ntena sement s edomnanty ydo ontann sxonseved ystene esdues a o w atate n dsude dn(22) and a vaae nume o otenta -ned osaton stes Fue6). Te Ctemna M sement esent n matue vons ontans a sotened etodoman ( amno ads and etans te two otenta memanesannn domans.

Te oaton o otenta ned yosaton stes AsnX-Se/T) oM s sown n ue 6 Seooa eated vuses sow oston onsevaton o tese stes Asn o te DN seoou o Asn o te seoou. Y ontans tee otenta stes wtn te eon and TB asne ste. A o tese stes dsa te sequene AsnX-T. Addtona



by




15/40


16/40


17/40

VIVIRU GNOM TRUTUR & XPRION 66

opio of u poi qu rv r of ii hoooy wl ivr (0 4) (iur ) h 2 y riu i hE coomi cy cov , i h c of W l ivov

i irmolculr iulfi bo () ( blowA mjor r v i h rivio of wo-imiol poiucurl mol Th mol um rlly cov rucur forvivi proi r b o rmiio of h iroluriulfi b for WN ( 22) b xmiio of opooil bioloipopi of piop fi by mooclol iboi (rviw i 68; lo 3 8 39 9 6 82) phyicl immuooiclhciio of E poi frm riv by pro or chmiclv (55 98) loczo of boy bii i o for

of h J E roi xpr i bcri (0 04) ucloi quciof urizio-cp u of B YF ( 8) rciviy ofippi wih iv fro V 6

Thr omis, sbili by isui bis wr oiilly fifm W E proi iulfi boi (22) Th iclu -milrio coii fiv oly hyophili cl rio, -i ro h hr rio r li by lor iulfifr rio hyrophobic chor i pr h rmiu Aumi h hi p of iulfi bri formio i rl fur of

flviviru proi h mol h frmwor for ioporiimmuoloicl ucur obi from umbr of ir viviu

h mo xiv ui hv uiliz h TBE poi n hou opo for TBE wi u hr Fur z dcoworkr (viw i 68 0) chrcri hr mjor piop clur (AB omi i h oomi by uiizi pl of moooliboi i ompiivbii y All hr omi oi piopivov i rizio hluiio ihibiio, bu oly oi

A oi flviviu ro-rciv piop Th xpri pprochouli bov hv low h omi o b orrl wih piulubrio o poi Doi A which i v from iu-rihrio of riu 025 h rio of riu 200250 (Fiur )coi vibl io wl hihly or qu (iu98) poibly ivolv i icyz fuio (0 ) vr urlzo-cp mu mp o hi omi for boh (98) (92)B oi A piop r iiv o urio by D ruci, or pi ih b xp fom h coy ucur o

hi om o uro pH-p oforio h) coi wih i poibl rol i ocyoic fuio v

Domi oi of typi-ri frm compii riu095 i ih for B oplx-pif piop h piop



by




18/40

666 AMR T A

are sensitive to reducing agents, but not SDS acid pH, or psin-eatment( Usng E suregions from J expressed n bacteia, the regon beweenresdues 096 has also een shown to contan both E-specifc an seo

goup cossreactve neutralzng etopes dependent upon a sulde crosslnage between resues 0 and 10 10 Two studies have povdedevdence for the bological mpoance of this doman n modulatngathogencty n vvo A neutralzaton-escape mutant of TBE that maps toTyr384 ( Figure 7 s aenuate n mce and confers potecton aganstlethal challenge cte n 70 Smlay a change at AP90 selected dungassagng of MVE n human SW-1 cels also appeas to lead to attenuato nmce (9 hese ata suggest that ts regon may be nvolve n tssueosm a erhas eect Ehost recetor teractos

TBE oma C s comose of a sufe-free loo an etoes n thsoman re ot eature y SDS-treatmet ut re rotease-sestve (he TBE gycosyato ste cosee amo avvruses mas to osto15 cose tre ure 7) wth ths oma 70 mato o thsgyca reers some oma C etoes sestve to DS enaturato 55

ermeta resuts ut atete sera or the V E rote ae general agreement wth the TB moe an cotute atoa shts(16) etes geerat atsera wth hhest eutraz actvty ma tothe Ntera reo suest that ths reo may e eose urther

more cometto eermets wth moocoa atoes a atserarase to etes rom sureos correso to the A a Bomas o B ect that these omas teract

The e o N-ke gycosyato ucto s ucer otetaN-e gycosyato stes e ot strcty coserve ut n numer anposton, smlates ae note amog seorou memers gure 6 A stenea position 10 is conseve among a DN seotypes some memers othe J suu a B ut ot Y Irect evece ests or theutlizaton of ths site n TBE (70 179 An addtonal ste at Asn67 s foun

for the DN goup contans a potential ste at Asn0 also found n SLEStes located nea the C temnus gre 6) ae poay not goo accetorsbecause of ther ocazaton n the proten hyrophoc oman (84either of the sequenced strans of W and KU vuses have redctedlned glycosylaton stes of the Asn-XSerTh type (28 172 t stherefore supsng that laee manose can be ncoporated nto ceassocated E but not the KUN extraceluar von E (18 Ths fndng mayreect strn fferences but coul also reect utlzation of otental AsnOXCys acceptor stes and consequent supton of disulde bond fomaton

Several studies examined favvus E glycosylaton n both veebrate andmosquto ce cultures (9 91 16 179 Evence exsts for oh smple andcomplex gycans (9 91 16 179 191 wh appaent host-secc df-



by




19/40

A M & 667

eene (BE (5 ) a we a eene epenng upon te tage o matuaton (0) Deene n poten gyoyaton ae a beenne o out Amean an Aan tan o YF () Gyoyaton and

eeae e JE en a been examne n eal 10 ewlyynee cellacae uced n ee mnkey kdney mqu(C/) e ontan a nge mannoe eno ente gyan onaoate E owy eeae no te utue meum (t ou) an te eno etane o etaeua onaoate omb e type nate tat gyan ae been onete to ompe m eleae eleae maed bu n able n e eence uncamcn ncan a E lycylan may be man meaue Fo TB on owee aboyate emoa oe nt

eue netty o aan e () atoug gyoyaton may t payome oe on aemby o eeae

Nal

N RTN Te lycoen ex a cel-acae (evewen 77) cell-uace 14 77) an exacellula nnvn 91 10 4) om Fau neton e antboe to N wt ompementn (CF) atty an te eete om o t poten a been ae teoube opemen ng (CF) angen ( Typeecc om

ex-pec an ueacve ee (evewed n 7) ave been ened an aea lay a le n ecve mmuny. Acve mmunan w ave mmunan w an mnclnal anbepotet anma agant aenge wt omoogou aue ( 4 4404 )

a a pete W o 4 wt atona moaton bylnke cylan Amn ac euence aa ncae a e emnu duced by naae cleavae (ue 4) an e C emnu bycleavae a a nvl oeae e lca ueam o te eaage te

ognay opoe o F N h poten nue ttyonee ytene eue (eept DN4) () naant gyoyatonte an atona egon o g equene omoogy A Cyp motcmmny un a e acve e l eae () lcae nea e C emnu alu n evdence ex an acae eaeacvy

ulle m o ae been etete an eene et betweenaue Fo an YF a poten ontanng N an a poton o NAan be ou wtn e ( 0 04) an etaeua u (0) A

puae N me a been etete ( wtn e an n exacelulaud DN and Pwaan T d cmlex can be dcae uce mnmec 1 by bln eamen w ac H bu n by D



by




20/40

668 CHAMBERS ET AL

(witut biin urea r reucin aent. Pible me ave abeen etecte r J an but tey aear t be iute uner me mcnitin (P an ena cmmuncatn I uylaet alle &

ce unuble ata. In uleae exerment te D2 meaea ate 2 mnute uetn tat ttanlatnal cen maybe mrtant mer rmatin (18

Te yica ierence between te membrane-aciate an ecreterm ae nt cear. embrane aciatn 1 a been nerrerm te ability eciic mnclnal antibie t lye viuinectecel in te reence cmlement (1 In aitin tw ecrete ave been rere; a lwy eimentin rm an a mre raily eimentin rm (9 2 1 38 8 Treatment J rm te cuture meium

wt nninc eterent w it eimentatn ate uetin blemembrane aciatin even ecrete rm (12 xeriment t examinete yrbic rerie te tein ave yiee cnlictin reult FrD2 newly ynteie mnmeric beave a a yriic rteinbecmin mre yrbic un imeriatin (80 In cnat celacate J 1 an 1 2 ( 1 beave me ke yrlc ten(1

arbyrate miicatin i imlie by te cnerve ate ine ycylatin ite l muitbe avivirue cntain tw

cnerve ite (iure 6 wit an aitinal ite in all Jcmlex memberexcet J A ierent atte eit TB wic are ne tecnerve te but a tw atial uniue ite Dect evience lycylatin cme rm everal tuie Cel-aciate D2 1cntan axmately u k -nke iaccarie wc cnt tructuraly etereneu manne-ric lycan rbably itibute vebt cnerve inke lycylatin ite ( 16 Bt en Henitive anreitant cabyrate a been etecte n D 1 exree by vaccinarecmbinant (191 r te celacate an extracelula an

' m mammaian cell an te cellaciate rm rm inect cellave tw -line liaccaie cain (0. r bt cell tye teiaccarie cellaciate are manne-rc extracellular rmrm mammalan cel cntan at leat ne cmlex ycan inicatin tatterminal aitin ccurre bere reeae ee lycrtein are releaelwly (t > m mammalan cell but are nt ecrete rm inectcel; ecretin i abie by nicamycin D2 rm mammalian aninect cel cntain tw ine liaccae cain; in cntrat t cel-aciate an extracelular rm cntain bt manneric ancmlex cabyrate ( 12 1 8 1 r TB viru bt te cellaciate an ecrete cntain uce an alacte uetin te reence cmex tye lycan (91.



by




21/40

AVIVRUS GNOM STRUCUR & XPRSSION 669

A a oe oorag he e aa or he NS 1 gooe o avabe a e reaiohi o eioe o he ia re oS s pooy undestood Antgenc domans have been dened at both the

and C-termn of E S ( 1 . Monocona anbody eptopes of DE2 havebeen ocazed o a reo bewee ao a 27336 tha contans a hghyconseved sequence (ree 32 to 33 a cystene-ch egon 129.

sA ROI The NS2A oei i he r o or reaive ahohobi roe (S2A NS2B SA SB o wihi e NS2a NS brego o e oroe A erie o roob reio areoerve o avivre (3 n oiio b o eee egha ee e are ebrae-aoae NS2A a bee iae e

oeig NS (ee above . The aare W o NS2A aer eraoa aa b SS PA ower ha he ree 225 . TheNea eee he UN proten Fgure 3ae a e eri of S2A may re r eavage b a ezeo ove sp:cfcty The ow C-eria eee o U S2A aea he roei o rae (re 3 Two or o he NS2A W2 a 22 ave bee oberve he rra bai or h heeroee inow (2

OI NS3 e eo are vra re pedcted W687 kd) s hghy conseved among favvuses (1 13 and contans noong hydophobc seches A oe fo S3 n va RA epcaton has beenposuated and ecent seuence comparsons (see beow) sugges that t soa a ea boa oag oh a oee av a oa eoie hohaae/heiae av. Thee roeie gge aoa: oazao or S3 wih ebrae aoao via ieraoa ave o bee ee

oario o he NS3 N-era rego (ree 18 w ear

eie roeae ha reveae igia hooog i or rego o S3 (36 (igre 8. A a Ser oie he ave aa ao he NS3 oeae ave e he oeve o XSerXPro o a NS3 aio a 111 a a oooo oo oher avivre reree he eee rog he eie oeaeeoh erie (6 The oeve A\ ree oowe b eiherTr Pe or Le or a avivire a he oerve eee -Ler--A- (NS ree 151156 or hoeie o o o he bae bg oe ha a eee eavage e

A e earer e vra reae a eave aer Arg bike r reer ibai ree suounded b amno acds wth shotsde chans Potease domans ae encoded by a numbe of postvestand



by




22/40

67 CMBRS

, , ,DN2 ( 4 3 ) GTHTVTRG [ l 3 a a ] ADVKKLIYGGG [ 4 3 a a ] DPTSVD KKG KWLYGNVE4 ( 4 3 ) EGFTMTRG [13aa ARMGGG [43aa TFTS RG GJ ( 4 3 VTTTRG [ 3 a a ] GVKRAGG [ 43aa] DRTS NG D GNV

KU ( 4 3 ) GVHTTTKG [ l 3a a ] GVKRCGG 4 3 a a ] TDTTSVD KNG DVGVME ( 4 3 ) EGTTTRG 1 3 a a ] GTGG 4 TS G GN ( 4 3 ) GVTTTG [ 13aa] GVKRCGG [ 4 3 a a ] TTTS NG GN ( 45 GGHTTRG [ 1 3 a a ] AVVAGG [ 44aa ] ATSN RG GN

TB ( 4 6 GVHTVTRG [ Ba a ADRCGGA [ 4 3 a a ] T AQG G

VDV ( 1 7 4 0 ) QGGVVTAG [ 26a a ] GVKTGCGAR [ 0a a ] DKNKSAG RVVRVKVKCV ( 6 5 0 ) QGG VVTCG 26a a ] GKTGCGAR [ 4 0 a a ] DKNKSAG VRVKK

HeCV ? NVWTVYA aa] YNVDNLVWPAPQ a a ] PSYKSCP A AVIFAAV

A ( 4 3 ) KNVVA [ KGGA [8 DGSVK GVGT ( 2 9) QDVTAACVG [ 34aa] NGTGAAQ [ 7 3 a a ] GDTCQSGRKDADQVVTR 3 2 SQWVVSAACYS [ STNNMS [ 7 ] KSCMGVVC SG QVSWSCT ( 3 4 VTAACGVT [ 34aa] TNTT [ 7 6 a a ] GCSGCG AT

A ( 3 7 QNVTCVR [ 37 VGYIAQ [ 7 8] VRGQDSGHVNG QAVVV

( 3 3 ) GMGT [ l l ] TAMEAQ 35aa] RGGGRSM G RAGA

GA ( 2 VAALTAGCT [ 12aa] TNGRHN [ 6a a ] NVCAQDSGA G TATGGB ( 2 TTAGCTG [ 1 a a ] GRT [ 61 ] CASG GT RATGA ( 2 TGTAGCGT [ 16aa] RVGAVT [ 63a a ] ACGRSGT AG QAQVMGN

Chymo # 7 102 1 2 1 0F 3 7 7 13 1 5 1

ge T NS poa doain Aignd unc o lavvu NS proin and cllulaproa urounding puaiv caayic id (w) of rin proa domain. Aoown a qunc fom wo piviu numd fom ginning of RF), ovin

via diara viu (BVV) (3) and og coa viu (HV) l0) paii viu (HV)5 and auopoa domain in mina gion of alpaviu capid poin foSindi (SIN) viu 59 135). Num f o dianc of f idu own from Nini of poin. Aviaion fo cua poa a pin (TRP); cmoypin(HT); aa (LA); Sepomes grses pin (SOT); Srepomes grses poaSOPA and SOPB) Sepomes es poa (SOP) sobe enzmogenes icpoa (AP) S x and Rfnc 3 and fo u dai

iue inluin n uttee min in te alaiu i

tein te etiiu te tyiu nuea inuin tein an teiai 3C an 2 pteae Te latte tee ae teine teaewit Cy elain e a te atie ie nulepile ee 3 an 6 eiew n eii eeene

3 l ntin inint ein mly wit te D---Dmily elie 7. Ti tte i eent in te mluptein te itie-tn iue an in ptein wit nuetietipate-TP binin mti Te euene alinment tee aiutein eneate een ein amin i neatin einte I

an III 4) Tee ement ae late between eiue 958 3 Ctemina t te 3 pee main. Bae n te mmnequene mti binin ite n B nwn TP-utiiin enymean utatie eiae ement I an II ae e t be tee ite te3 ptein ement I ntain te nee mti l-X---Te



by




23/40

FAVV NOM TUCTU & XPION 671

(ree 15 for a eget II the otf AlAaA(ree 99 Aother hghl oerve rego, orreog to eget I (e 5 a be a le a bg ste (7 Seget IA a III are es oserve rego of ow fto Noevee ort leavage of the NS3 rote to earate roteae aheae oas

T N N N N N hese sa rotes (gre 5le NS are oor oeve aog avve ( 1, bt otasar sttra featres osstg reoat of tle hrohobteta ebrae-sa as The NS2B a NSB tes aereal etfable fete els, bt NSA has bee eftve etfe

for UN (9. No sttraslatoa ofats of these rotes areow este the resee of a osere, otetal N-le glosato h mnl pn f NB he pn a fmebrae ets f the vral reat ees a be vove ebrae loalzato of NS3 a NS5 va rterte terats

NS PN NS5 the larget (ete MW 131 a othghl oerve of the avvrs rtes ( Cteral seee atafor KUN ate that the rote ees to the rete e of the OR( 1 NS5 a bas rote a e ot ota a log hrohobstrethe The osble role of NS5 a the vral RNA olerase see bewa the geerat ts tes b eavage the tas oartet (reabl b NS3 or a ateatve rotease gget that NS5 soate e tola alhogh t aoate wth ebrae

Rego of eqee oevato are trbte throghot NS5 A hghloeve oa NS5 ( ree 53 to 75 ota the eqeeotf -A-A (133 13 whh reet otrtral rotefro a ber o otvestra RNA vre a beeve to a a role RNAeeet RNA sthe ( 7 13

VIRION STRTRE N SSEMBLY

lavvrs vros ota a leoas (53 aeter srroe ba blaer erve fro host ebraes otag the eveoe rotes a M rM The aeter f the vro aroatel wth

sfae ets f 5 ( 1 3 Althogh holgal stgshable tw vro ors have bee haraterze traelllar vrs ota ol rM whereas etraellar vos release to theltre fl ta reatl M (fr revew see 13 (gre 9



by




24/40

67 HAMRS A

Inraclllar ron Exaellular ron

Nulepd

gure 9 Srucure of inracelular and extacar avivis virions

Ulsrl ss h ro mophognss ors sson w nacelar membranes (revewe n 3) Elecrn mcrscopcss of avvrs-nfece verebae (78 89 06 1 3) an msq (35

65 8 1 16 1 50) el cres as wl s n o ss mose brn 1 5 1 ) he onssel obsee mohoogcl mr os rs whnh lm o h ER. I m ss, os ppr o mle whnsorerly arrays f membranebn vescles. Bng nemeaes anceary sngsable cyplasmc nclecapss ave n been fenbsrv sggesng a e assembly prcess ccrs raply Dramaceron o e nre membes s m o s neon he spo o s os, om he ER o he l se whrxooss ors s b o nolv escles h may arse from ER

(35) or hr compnens f e s secreory sysem (65 50) Aleave, ssoon of the vescles may ampany cytlyss g e sgs offo Bng f vrons hog h plasma membrn hs beeobs o nree (6 106 1 150) os o ppe o b mjor mehsm or on reese

Ts obsvos oger wh h scr formaon avalabe or on pos n er moe o syess (121) sgges mol orvrn assembly an maran e cyopasmc gly basc man f heC pon prsmbl nrs wh va geomc RNA o o ops precrsor. e ropobc segen o he achore C poe m seeo ocl ssmby o cocapss o memb ss (21), lsofnon as an nea sgnal seqence or prM (10) Trnsoaon o heprM E pros posons her coomas wh h lm o h ER her hrophob C rm lp ber. Ths oon o CprM an E w respec o he ER embrane sggess h 1opsscre eeope by bng o he lmn he moll rosnvove hs ssmb pocss e no be s b E prM s

prse raellar an extracellar vs ppar o b ssoc seergensable eemers (74 Bh prM an E are pre o hveony small cypasmc mans, b nlke smple rsmembr nchorsh hyrphob C n re hghly onsere a col be oe nE-prM or opncops nros Ths moe prs h h



by




25/40

AVIVIRUS GNOM STRUTUR & XPRSSION 673

anchored ceavage occurs on the cytpasmc side of the R. In view of heconseed basc amno acd par preceding the vrion -anchored ceavageste ceavae ma be medated y a ra teae eha N N na

tn eucdate te tmn anced C ceaae wc may ccu mmedatey te anatn ate dun rmatn te nucecad.dcatn (r me re and r ycan y mmn and

temna addton mpies movement of newy formed viions tough apathway smar to that used for snthess of host pasma membrane gcopotens 2 2 2 ). A roe for N S n vron assemb tanspo or eeaea ed t RA ecatn a een ueted ecaue t ycyated and eceted m.

Te deaed ceaae (ee and e lumna ocazaton of te

ceavage sie suggest that the event may occur n a comprtment of thesecretory pathway medated b a host potease or possib a vira protease

(34). Te ceaae te equene A--Ary-Ar (whee X vabe) ma tat ecned n matuatn ceaae numeu aycten (eewed n ) and ceu -ten Te naty tdetect ntaceua cntann on uet that th ceavae occujut por to or durng reease of vrion. The boogica oe for prM ceavageis uncear but ke smar gycopoten ceavages for other enveoped vses) t ma trger changes n the von enveope that promote nfectivt

In t ead ne tudy ( eed tat the ecc nectty WNntaceua u 6d we tn extaceua u [wc a aecc nctty 6 ace e aquermn unt P]. Areorganzaton of the WN viron enveope a een reed t ccu wtpr ceavage because detergent-stabl - hetedmers ae not oservedbut nstead apparenty trimerc compexes of the E proten ae seen 475 In cntat t WN cnn deteent-ued extaceuTB eut n eerent matn ten dme (69. Atu teeeut uet ac derence n enee tuctue etween mqut

boe and tboe avvruses, furer stuctura studies ae needed beforem concuons can be made

SD SQUS D STUTUS

Sho conseved RA sequences and compe secondy ucure ayimpotant acting roes n transatona reguation as we as n Arepicaton nd encapsdaton of a number of pant and anima RNA vuses

ompasons of avvs seuences have ed to the dentfication of eveatentay mnt eement. ue w a cematc eeentntee cneed RNA equence and uctue tat dcued ew. Teunctn n recatn if any remain speUative and need t e



by




26/40

CHAMBRS T AL

5'

_

-

_

O

Suctral Proeins

osural Prens

3'

5S

DEN2 DEN4

3 NC \S SI

.

-.I

10 Cd d rpd RNA q A t g t dg 5 C t p dg OR) dg t ttpt d tt prt d t ' dg gi (' C ditd S bd d t t d dg g l ggtgrvd 5' CS CS CS2 d rpd iry ld bx q dbd ttt AUG d r t d g t OR dtd by t b r b

t py A dtd t rt OR t 5 ' dg g BE.Sq r btd rm t r gi t gd igr

ar by xprimnaion. Non of N faur ar by nom of mouio-bo avivir av bn inifi in TB.

an 3 ' mna cn tuctusSconary cur con n conformaion an abiliy bu no in

pimy unc can b pric or 3 ' inal 90 ba o mouiobo vi , 50, 58, 57, 60, 7 , 90). T pricabiliy G of i aipin loop rcr vari in iffn flavivirfrom 0 o 0 cal/mol. Svral lin of vinc (for rviw ;alo , 5 8 , 0) uppor xinc of uc conary rcr . Sm-loopcur av alo bn ona ily a 3 ini of vralaii o plan vi cid in 58, 5). For o vi uc curar uba or amino acylaion in vio an in vivo an av bn implica in iniiaion of minu-ran N yni Similar ui av no

bn rpo for flavivir.lou abl mloop rcur wr no iniially pric for

WN 5 ' -rmnal rion (), Brinon & Dipoo 0) inifi ponialcondy cur vau ro 7 o -28 Kcao n 5 '



by




27/40

AVIVRUS GNOM SRU & PN 67

rmn o a ar gnom nng D2 D WN an wit te oponng po n te n ominu-tran .

Cvd ad Rad Sq

Two o conerve NA equence calle C an C2 iure 1are by all moquito-boe avivirue are locate ' to te putative 3'terminal conary tructure 8. C i about 26 nucleotie in lent anocate immeiately aacent to te terinal econar tructure. P of C 1i compementary to a conerve equence near te en of te enome inte reion encoin te capi protein ' C Bae-pairin of tee or oterterminal equence coul lea to cyciation of of te viral enome tat may

be importnt for reulatin enome traation replication or packain 8 17 Wile evience upport cyliation of apaviru an bunyaviruRNA ( n 58) mporan o n paon unclear. Cycliation of flaviviru RNA a not been examine Te motily conerve equence C2 i approimately 2 nucleotie in entan i locte 1 2 to 22 bae ' to C C2 i uplicate in member of teJE an DEN ubroup iure

Beie tee ort equence tat are conerve amon flaviviru ubroup broup-pecific feature are alo oberve a a et of tree

cloely pce repeat foun between ucleotie 137 an 12 33Tee repeat eac about nucleotie in lent contain four to ixierence in pairwie comparion. Member of te JE uboup ave aifferent equence about 2 nucleotie in ent preent in two copie inte 3 nooin reion In ontrat to tee p are eparate by 1 2to 13 ba In te noncoin gon W contain tee ort equenceix to even bae in lent repeate oce . ember of te JE ubroupao ow inificant nucleotie equence omoloy in teir ' noncoinreion TBE contain a uplicate equence of about 2 baeowa te 3 ' en of te enomic RNA; one of te repeate equence i in teregion nong te C min o an o n ' nononggon 1 I ill o n o ompar qn o mo gntic-boe avivirue to ee if oter conerve NA feature can be ientifie.

RA REPLIATIO

After tranation of te incomin enomic mNA RN replication invoveyntei of complementary minu tran tat are ten ue a template forprouctio of aitiona enomelent plu-trane molecule Tee putran cn ten be ue for ranation of rcura an nontructural



by




28/40


29/40

AIIRU GNOM TRUTUR & XPRION 677

tobacco moaic vu the domain of the nontuctua ptein containng theconeved Gy-Ap-Ap polymeae motif 134) ae pduced in maquantitie by eadthough of n-fame teminaton codon.

efetive-intefeng I) paticle e vauable tool fo the tudy of NAvu epcation nd my play le in vi pathogene n ome ytemee below). o ome vue tongly ntefeng ptile ae eygeneated by hgh mutplcity page 16) nd ontn unted and eanged genomi NA. Thee NA ontan -actng equence neeayfo epltion and pakaging but uualy do not encode v poten. Theyheefoe need a helpe vu to upp thee funtion in In avviue ptce hve been obeed n petently nfected vetebte celcutue but tongly ntefeng I ae not eady obtaned unde thee

condton o duing eil hgh multplty pagng . eveal pobltie my expain thi obevaton nudng the hypothe tht unde hecondton teted, mot of the viupeced omponent of the eplatonmchne e eue n cis oev compementton t lo leel been demontted beteen tempetue-entve mutnt of J 40) o L74) tht ave defet n NA ynthe.

A ytem n whh ppea to be eadly geneted ha been tuded nome detil by Binton nd owoke ee fo eview). tudie that beganwith the oevtion of hetble ueptibility to Y viu in mie 1 39) ed to

the dcovey that a ngle utooma dominant allele n onfe eitance toavvu nfeton ee fo evew). lvvue cn epcte n etntmce poeng th lele but the ped of WN nfecton loe ndgnfnly lowe peak vema e found 0- to 10-fold) than n ongenueptibe mice WN eplication ha lo been tuded in pimay iboblatfom thee tain n cel fm etnt ice va NA ynthei waeduced owe tite of nfetiou viion wee eleed and a hgh popoton of I w found even fte a ingle gowth cyle Thee eut indiatethat a pecc but a yet unidentifed hot gene cn hve damtc eect on

aviviu NA ynthei The identifiaton of thi hot gene pduct nd thecel type telf in which defective fiviu NA can be eadily genetedand ppgated could have temendo value in futue tudie to deteinethe -acting equene invoved in NA eplication and packging.

MPRT SPTS

ntic Divrnc amon aviviruss

Claifiion of flaviviue ha titionally eied on elogia tet.

Amno dequene data n now e ued to compae the eatedne ofdffeent flvivue and the eult of uh ompon e in genegeement wth thoe deved om c-neutaation tet with polyona

Annu.

Rev.Microbiol.1990.44:649688.Downloadedfromwww.annua

lreviews.org

byUniversityofMichigan-AnnA



30/40

678 CHAMBS T A

2

3

D4

49 45

4 47 45 47

4 4 3 37 43 4

36 35 36 36 36 3 3 3

Z "

z

fz

z Q Q Q ; ue 1 1 Homology matrx betwen diffrn avivirus poteins Pirwise compaisons ofligned sequences (% identic residues) Seologic subgoup clusters are inicated bydelined old tye

nisea (4 , 100 1 24 14, 166 . r 1 ar E rseuene hoology (% enl eses eween seel een liuses om hese aa he subgo elaoshis ae aaen he DENseoyes cluse s he membe of he E subgou is isly

elae o r TBE w lar vr fom a of qb vr r b aa ad S qsuggess ha E is somewha close o (100.

Oligonuleoie r olymohsms av beeuse as eemiological ools o suy he vr of ieeenisolaes of ula ises iein n he geohi loaio me ofisolo imal r a a r vrl henoye. uieswi DE ( 0, DE2 ( 1 6, 1 64, DE4 (2, ME ( , 6 SE ( 1 6,n (6 eveale a emaable genei homogeeiy alough siffeeiion c b eece in nal coeles wih geoghicoigin oe eely lime nuleoie seuene oson of VE (4DE (2, a DE2 (6 isoles yiele smila esuls. uh of hegenei sabily obseve fo avvr i naue obbly ees hei hosaaa a aa alav y

Ra a r, r w aln liil a o sese seveiy mgh allow he efiniio of sei arkr r vl T ar lar r has Anohe oh o hs uesio involvs he comiso of aeuae vccinesns a he iulen aens. The seuenes of aeuae sis of 57) a JE ( 120 av b ar r vrl aral a. Bohenue sins ife o only i he vrl mes bu also in hiably b rad by q ar d a br



by




31/40

VVRS GNOM STRTR & XPRSSON 79

nuceide ad ain acid chaes disibued huhu he enes. Fexale, he YF 7D vaccie sai nd is vile aea sain, Asibi,which wee seaaed by abu 24 seial assaes, diffeed a 67 ucleide

siins (7, 132), eading 32 ai acid subsiuis. Give he abiiy ecve infecius virus fm ce cNA (13), i shu w essible eerine which of hese changes ae ilicaly ignican

mares o Oher Pose-rand Vruses

Cmaisns f ucleide an ami aci sequences (eviewe i 13, 1)an i roluion rucur minion of anima an lan viru aveveae cmmn hemes as well as a ivese secm enme ganizains and sraegies f elicai. A discusse abve (see als 3), virus

secie einases are cm any siivesan virus amiies asare sequence mifs suesive f NPiig mains, helicase ezymes,ad NAeeden NA lyease aciviies Alhuh avivises weeiginaly gue wih ahaviuses bse hei siila hlies anes f asissi, i es f gee anizai an exessin, hefavivises cleary use a saegy simil he ica-ike viuses 88) Bhvia ees ece a singe lyei ha beis wih he srucualreis a is elyic ally cesse yied he aue via eins. cas, iri heesis an scua ei cessin f avivi

uses ae sy siila bella vis (42, 73) which akes u a seaaeeus n he avis faily alng wih he alhavises (see as 43) Bhviruses bably uiie eane-ached ecus rs f he C reian acquie i eveles y uing hruh iaceluar memanes.Ms clsey eaed he avivises hweve are he esivirses, as aeus f he avirus faily ece gee sequences f bvie vialiahea vius (BVDV) () ad h chlea virs (HCV) 08) sues asimia gne aanee and elcai sraey aviviuses (29)Hweve scan ain acisequece ly exiss beween he esivi

uses an he laviviruses, a hmgy is ailfud i he ifsicu abov for NS3 a NS he ecey ienified age f nn-A,non-B bloobo via heaiis (heaiis C vius) (22) als shaes siialimi omoloi wih he faviviru an esiviruses (C. ice & .Feinse" uublished aa) (Fiue 8).

OUDIG RMARKS AD UTUR PROSPTS

ocul ui on faviviu av enee an loiv a. he avra yar av n an amain accumuaion of rucural informaionan common a w a iinc faur of aviviru nom anroin av bun o mr. Howvr for nary vry of viruif cyc maor a in our unranin il i rly vn in viru



by




32/40

8 CHAMS AL

ost ntracton suc as rctor bnng ntraton an uncoatng ar stoor unrstoo. Substanta rogrss as bn ma n unrstanng raorotn rocssng. n contrast t s knon o t mocuar n

tractons morant or on ormaton an t assmb an uncton o tmmbran-assocat N rcaton coms . Th constructon o cDNcons caab o roucng nctous N anscrts 12) shoul actata rct molcular gntc aroa or stung man o ths aras oarus bolog . St-rct mutagnss an consucton o rcombnantruss t omoogous or troogous substtutons l allo sucturuncton analss o iral rotns an NA squncs an gntc mango an ntrstng boogca not nclung cangs morant oratnuaton n rbrat hosts an or hroo ctor comtnc. unc

tonal cDNA clons can aso b us or rsson o nual raotns or roucton o N subsats or omnt o n onmatc or N bnng assas. Man otr anus ar ort ursung.mong ts t omnt o n tro N rcaton sstms caab outng ognous tmlat Ns ou b nauab or stung arnts n N rcaton. Ultmatl a hg rsouton tr-mnsonastructur o a arus E rotn an/or on b u n unrstanng ron assmb an rus-ost ntractons On os that contnungaancs n our basc knog o arus boog l ntua a to

mor ct mtos or conong ts mortant grou o atogns.

ACNOWLEDMENTS

W tank . Dagao S . onon an M MacDona or man ucomnts on t manuscrt C. Man Hn an D Trnt or rongmanuscrts ror to ubcaton P Mmora Trust USAMD tockr ounaton an NH or nanca suot. C S . Han s aohnson an ohnson lo o th W aoo to o colas o omsson o cran subcts an th ncomlt traur ctatons bcaus

o sac mtatons.

L

1 . Baltmor, D. 9 Srucure of thepolovus replcatve ntermedateRN J. o 29

2 B J F., eeick, 988 . Viral cytene pteae homologou tote tpnle famly of ene poteae: Strucual an functonal mpton. U

7727 Bazan J. F eterc R 99 Detecton of a ypn-le erne proteaeoman n flav ve a p veVroloy 1779

Bell, J. R Knney, R rnt D.W Lenche E Dalgo L,Sau, H 9 Ntemnal amnoaci equence of ctura protein ofhee avivie Vroloy 2229

Bezyca , Cauc R, Baolomeuz Gorma Wrgt P 8 hteton of potee

cleavae te nvolve n the formatonof the envelo glycoproten an reenonrucural poten of engue vtyp 2 New Gunea C an J GnVrol 72



by




33/40


34/40


35/40

FAVVRUS GNOM SRUUR & XRSSON 683

4 Gould A, Buckley, CammackN. Bae A. D. T. Clegg J. C. S. etal 1 985. Examination of the immunoloial rlationships beteen lai

vue ung yellow feve v monoclonal antibodie. Gen Virl66 13698

0 Grange Bouloy M Giad M18. Stale econdy tcue at the end of the genome of yellow feveis 17D vaine stain) FBS Let.1881596

1 Genbrg , Woo, W 5 . , Biedryca Wrght J 988 Patialnuleotie sequene and dedued ainoaid seuence of the stctual pteno dengue vu type 2 ew Gunea C

nd O18 an 1 Gn Vir:13152. Gn, B., Bnton M. A. 18.

Chaact{aton of Wet Nile viuNdepnt N lymase ancelua tena adnyy and udyytrnsfese in cell-free extat. Virl.0 1 1 1 32

3 Gn J B Bnton M 987 Dissociatio of NS fom ell fractons ontag West Nile vis-specicpolymee ativity J Vro 61 3-

4 Grun J B Bnton, M. A. 1988. Separaton of functonal West Nle srepliation omplexe from ntracellularmembae fragments. 1 Gen. Virol9212

Guirakho, F , Hen, F X Kun, C989 Eptope model of tik-boe encephalti vs envelope glycoprotei Enalyis stutural propeies role ofcbohydte de chain ad confoational change occurng at acidicpH Vr 9099

6. Gukhoo, Radda C. Hein FX Kun C. 19 vdence for anti

genc talty of tk -boe encephaltivrs by e analysis of natural isolates en rol. 8854

7. Hahn, C. Dalmp . M. SaussJ H Ri(;e C M 187 Comparon ofthe viult Asibi rain of yellow fevervis with the 17D vane stain devedfom it Po Natl. Aad. Si. USA84:013

8. Hahn C S Hahn Y 5 , Rice C MLee, E. algo, L . , et al 1987. Coned ment in the 3 untranslatedegon of lavv RNA and potential

cylizatio sequence. Mol. Bio.1983319. Hahn C S . Saus E. G . Sauss J.

H 98 Sequence aalyi of teeSndbis viru mutant temperae-

enitive in the capd pten autoprotease Pro Natl. Aad. Si. USA8:46485

0 Hah Y S Gae, R. Huapier,

T., Dalymple J M., Stau J H.Staus . G. 1988. ucleotide equene of dengue 2 RNA and ompaion of the enoded potei th thoe ofoth avv Virolo 121780

1 altead B 1984 Seectve pmyhealth ae Sateges for onol of diseae n the developing world XI Dengue. Rv. Inj Di. 6164

Haltead S B 1988 Pathogenei ofdengue Challenges to mleulr bology. Sien 23947681

3 Hae . Summer P ckels K

H 1989 lavivr ent nto ulredmoquito cells and human periphealblood monocyte. Arh. Virol. 1041243

Hase ummers L Eels K.H., Bae W B 187 n elecron andimmunoelecton micocopic dy ofdengue- rs infetio of ulturedmosuito ells maturato events ArhVirol. 31

65. He Summers . L., Eckels K. ae W B 98. Maturation process of Japanese enephltis vis incultured moquito ells in itr andmouse brain cells in io. Arh Virl.961355

6 Hahimoto H Nomoto , WatanabeK . Mo T Takezawa, T et a 1988.Molecula cloning and complete nucleotide equence of the genome ofpnese enephlti is ejin- strain Viru ene 130517

Hawkes R A Roehg J T, HunA. Mooe, G. A. 1988 Antgenistture of the Muay Valley enephalitis vis glyopoten. Gen Virol1 0

8 Heinz . 1986. Epitope mapping offlavi glycopoteins d Vir Re311038

69 Hen . . Kun C. 10 Ioationof dimerc glycoprotein ubunit fomtick-boe encephalit vir. nterirolo 13:1977

. Heinz F. X . Mandl c Holzman H.,Guiakhoo F , Tuma, W. , Kunz C1990 he stue ad ution of the lais enelope proten E.n nd nteationl Spoiu onPotie Strand Viru, Venna ed M.

A. Brnton F. X. Heinz WashingtonD: Am S. Miobiol. n pess7 . Henhal A., Hehal L. S.

Thaiomboonuk B. K. 19 Topologcal mapping of unique epitopes on he

Annu.Rev.Microbiol.1990.44:649688.

Downloadedfromwww.annualreviews.org

byU

niversityofMichigan-AnnArboron02/26/11.

Forpersonaluseonly.


36/40

68 CHAMERS E A

denue-2 viru NSI potein uinmonoconal antibodie J Gen Vl68:84551

72 Hencha E A Rep, P M,

McCown M , Brandt W. E. 1986.Identifcation of an tigeni and geneivrant of denue4 vrus from theCaribbean. A J Top Med H3:4010

73. oban T C Gim S 1989. nv and n vv expreion of beavi gycopoein 2 Te signa pep-ide i s ontained n the Cterminal regionof apsid proein. Volo 17241

74. Hoinhead G Brawner T A emin T. 198. S. uis enep-iti vir temperareeniive utans

I. Induction, ioaion and preiinaychaactezation Ah Vl 751717975. Houghton, M hoo Q. , o G

1988 u Pn Applon No88310922.5 Publiation number0318216

76. Inokuhi Hiahia A 1987. I-teerence wit vira infection y de-fetve NA repliase J Vrl619469

77. ie K Mohan, P M aaui Y . Putna R. Padanabhan 1989. e-uen nalyss of loned dengu virstype 2 genome New GuineaC strai.

Gene 719721178. Ihak R Tovey D G owad C R

1988. Morphoenesis of yelow fevervi 17D n nfected el culures. Gen Vol 69:32535

79 amer G ro P 984 mstuura omparson o Ndependent polymerass om plan n-ima and bateal vuses. NulAd e 12726982

80. Kb K 1980 Gene chctsts and ntgen relatonshps. eeRef. 108a pp 10558

8 1 . Ku T Oha, A 1988ociation between the pHdependentonformatonal hnge o West Nie a-vivirus protein and viusmediaedmembrne fusion. J Gen Vol69 1 2474

82. ma-uda J. Yasu K. 1986.Antienic comparion of enveope potein E between Japnee encephaivu and ome other avivue unmonoclonal antibodes. J Gen Vl6726672

8. Ko K. K. grash, , ukai, K979. eton mosopi oeatonon Ades alboptus e infeted withdenue viue Ah Vl 62412

84. Kofeld R , Kofed S 198.Aembly f asparagnenked gosa-

chaide Annu ev he 546164

8. Koak, . 1986. oin mutation definea equence ankin the UG initiator

codon tht moduate tanation by eucarotic iboome Cell 44:289286. Koza . 1989. The snnn model

for rnsation updae. 10822941

87. ruich H G Wimme . 1988.ral protenae Annu ev Bhe577014

88 Kuhn R J . Wier E 198. herepicaion of picoavie In heMoleula olo /the Pstve andVr ed. D. J owlands M. Mayo B W Mahy pp 175\. Lon

don: Acadeic89. Leay K Br C D 1980. Sequentiaevent in the ophoenei of apaneeencephalitis vius. Ulasut es72 12329

Lee ., eon C., Sipon R.Wir, R c, Rice C. M Dao L1990. equence of the 3 I haf of heMuray Vaey encephaiti vugenome nd mappng of the n-tuctu poein NS 1 NS3 nd NSVus Genes In pre

e, M Crooks, J tephenson,J . 1989. The synthess nd matura-

tion of a noncra extraceuaranen om tiboe enephaitsviu nd t retonhip to the ntaelua N1 poein Gen Vl7033543

9. Lobigs M, Dalgo, L Schesnger J Wr . C. 1987. ocaon of aneutralizaion determnant in the pro-ten of yelow fever vs (I7D vaneain . Volo 161:47478

93. Lobi M. ashall . D Wer .c Dg L 1986 Genetic differentiation of uay alley encephai-

ti in uraia and Papua New GuineaAus J p o Med 64718594 i M, ashall I D, W R

c , Dalo L 1988. Muray aeyenephs vis fed sains om utraia ad apua Nw Guinea: Stdie onhe seuence of the major envelop pro-tein ee and vence for ice Vlo 165:24555

95. Lobi M Uha, R Netoowc AMarhl I D Wr R. DgL 1990 ost el selection of uayey enephaliis virus vaiants edat RGD equnc i the velpprotein and in moue Vol1765879

96. Loi M. Weir R c , Dalgao L1 986 Geneic nass of unjin io



by




37/40

A VIVIRUS GNOM STRUCTUR XPRSION 685

lates ng III ad I esondgests of sngle-sanded NA to von A. A Ep B Md S6418596

97 Mackow E Makno Y Zhao, B Zhang Y M Marko, L , et al.1987 e nucleotde sequence of denge ty! 4 vus Analysis of genes odng for nonstrctural poens. Vlgy59:21-28

98. andl, W., Guirakhoo, F. Holzman, H. ein, un, 1989.nigeni uue of he avivus envelope potein at he molecla level,usng tkoe encephalitis vrs as amodel. Vl

99. Mandl, C. W. , en, . Kn, C188. S'qence o the strctural poteins

of tc-boe encepalits vrs (Westesbtype) and comparaive analyss withother flavivuses. 16619720

z tc, Knz, C. 989. Genom seqence otckboe encephltis vis (Wesesbtype} nd compaatve analysis ofnnsl pen w ohe avvirses. Vlgy 73930

0 1 Mkoff 1989 n iro pesng ofdengue virus stcta proteins cleavage of e pre-mebrane proen

Vl 635-52102. ason, P. W. 1989. Maturation ofapane enephalitis vis glyopoteins pdued by ineted mammalianand moqio cells. Vlg 1693546

103 ason, P W . , arympe, . , en M. McCown, J M, Hoke C.H et al 189 oleclar chaacteriation of a neutalizing domain o theJapanese encepals vs stuaglyoprotein n Vl 7020749

104. Mason, W., McAda, P. c Dalrym

ple J. M Fouier, M. J Mason, T.L. 987. xpeo of apaee encephal vrs ntigen Ehhacl Vlg 158:361-72

05. Maon, W MAda P. C. MaonT L. Fe, M. J. 987. equeneof the dengue-l vs genome n the egion encodng e three srucal proteins and he major nonsctal poenNS Vlgy 6:66

106 Masuma T. iraki K. asitkaT Hot, S 1977. ophogenesis ofdenge vius in lues of a humanlekemic leukocye lne (J- l l) . Mc- 1 3934

107. McAda, C Mason, P. W., chmalohn, C. S., Dalymple, J. M., Mason,T L. oier M. J. 987 Paral n-

1eode sequence of e Japnese ncephalis virs genome. Vlgy58:348-60

08 Meyers, G, Rumenapf, T hiel, H. J.1989. Molecla clonng and nucleotde

seqence of the genome of hog holeravs. Vlgy 1715557

108a onah T . , ed. 1980. uEncpalts Washngton DC: AmPublic Health ssoc.

109. Monath, T 1 985. Glad tidings fromyelow fever esearh. cnc 2297-35

1 10 Monath, T 1 986. athobiology of theavvises. See Ref 144 pp. 375440

1 1 . Monath, T 1987. Yellow fever: Amedially negleed dsease Repo on asemna. R nct 9:6575

. Monath, T P. 988. apnese enephaliisa plage of the Oent. w EnglJ M 913

1 12a. Monat T P 990 Favv. InVlgy ed. B N. Felds, D. M.Knpe, pp. 7638. Ne Yk: Rven

1 13 Muphy, . A 1980 ogaviumorphology and mopogeness. SeeRef 3, pp. 436

4 Mugay M. nmn P. J Hozinek M C Weila E 1975.Growt cycle o abovise in vetebate and atropod ells g M

Vl 958323 Nagamatsu . ikch grashi,A. 1988. emnal seqenes of he epliave fom of of the Japaneseencephaliis vs. Acta Vl (aa32:75-78

g, M . 98 . Utrasctural stdiesof Kunn vius-nfected A s cells Gn Vl 687782

7. Ng M. L., Coer, L. C. 989. Detecon o some denge- angens ninfected ells sing immno-miosopy.c Vl 04 9708

1 8 Ng M. L. , Hong, . . 1989. Favvrsfecto eea laalchange and association of Knn vsNS3 protein wth mcrotubules. AcVl 106 00

9. Ng, M. L, Lau L. C. 988 Possbleinlveme o eept n the en ofKnjn vs into Ve ells. AcVl 00:992ll

0 Ntyaph S Gant A Trn DW . 990. Te ull-lengt nucleotde seqence of the vilent A14 sain ofapanese enephaliis vius and itsattenuated vaccne derivatve, SA-141. Vlgy n pess

12 1 . Nowak, T arber, ., WenglerG., Wengle, G. 1989 nalyes of hetermna seqenes of West Nle vrus

Annu.R

ev.Microbiol.1990.44:649688.Downloadedfromwww.annualreviews.org

byUniversityofMichigan-AnnAr

boron02/26/11.

Forpersonalus

eonly.


38/40

686 CHAMBE E A

structural proteins and of the in rtranslaio of hese proeis allow thproposa of a complee scheme of thoteoly cleavages involved in thersynhess. Virlgy 169:36576

22. Now T., Wengler . 987 Anayssof disuldes present in the membrane

poteins of the es Nile flavivirs.Virlgy 156: 12737

123. Ohyama, A., to, T., Tanimura, E.Huang, S -C., Hsue, Y., Furu Y.1977. Elecon microscop obseationof he budding maturation of group Barboviruses. irbil. mmun.2153538

124. Osatomi, K . Fuke, Tsu D. hba T Sakaki Y. Sumiyoshi H 1988Nucleotid sequence of dengue type 3vus genomc A encoding viralstrctural protein. Vrus enes 2908

12. Oden S. oirer B. 1985. enguevirs nduced polypepide synthes.Arh. . 85: 12937

126. Perraul 1981. Ogn nd repicationof defectve nterfering particles. Currop o mmo 9207

G, Yamshcikov V .Blnov V M 1986 Tick-boe ecephaliis genome: The nucleoide s

quence coding for viron strucural proeins. FBS e. 200372127a. Plenev, A. . Ymshchkov V

Bnov V. M. 1990. Nuceode sequence of the genome and complee amno acd squence of he polypoen ofick-boe encephais vis. Vrly17463

128. orerield, J. S. 1980. Antigenic chracterstics and classificaon of gvrie. See e. 13 pp. 36

29. Pu J R., Ches, P C., Padmanabha . re K Hoke . H,

Burke, . S. 1988. uncional and antigenic domains of the dengue-2 vsonsuual glyopoe N. oogy 1639303

130. Repik, . M. Dalympe, J .Brandt, E , Mcown . M., ussel, P. 983. RNA ngepining as mehd fo dstingushng denge virus strans. Am p. e g.3257789

131. Rice, C. M. Aebersold, R., Teplow, D.B Paa . Bell, J R e l 986Partia N-erminal amino acid sequenesof three onstructural protens of

twoavivises. Vrlgy 5 932 Rice C. M., Goui A. ller R,

Chambs T 1989. Transcrpion ofinfectious yelow fever vs RNA fom

flllengh cDNA tempaes produced byin vo ligaio. New B 128596

133. ce C. M. enches E. M. Eddy S.. Shin S J Shees . . StraussJ H 985. Nucleoide sequence of yellw feve us mplias fo favirs gene expression and evoluton.Siene 229:72-33

134. ice . M , Sauss E. G Strauss, JH 1986 trcture of the avvirsgenome. See Ref. 144, pp. 279326

35 Rce C. M. Strauss J H 1981. Nucleotde sequence of he 26S A ofSndbis virus and deduced sequence ofthe encoded vius s poteins.Pr Nl Ad S SA 782062-6

136. oerig . T., unt A. ohnsonA. J Hawkes, R. A 989. Synthetic

peptides derved from he deduced amino acid sequence of the Eglycoproeinof uay Valey encephaits virs elicit antiviral antibody Virgy 171:960

37. RuiLnes A , Chou A. , Despes,P. ird M. Bouloy M 1989 Processng of yelow fever vrus polyproten: Role of cellul oee imauraton of he struca poteins. JVr 639209

18. ussell P. K Brdt . E alrympe M. 1980. Chemical and anigenc

structure of favivises. See Ref. 143,pp. 50299 Sawye W A loyd W. 1931. The

use of mce in tests of immuniy againstyelow fever. J xp. Med. 545355

40. hlesinge, J. Badss M. W.,Copp, C. B. Monah, T P. 1986 Poecion against yellow fever in monkeysby immunzaton with yellow fever virusnonstctural potein . J Virl.60: 5355

41 chlesnger J . Bradrss M. .,Walsh, E. E. 985. Protecion agains

17 yellow fever encephaits n mice bypassive sfe monlon nibodies he sul glyooegp48 d by actve immuniation withg48 mmunl 3528059

142 Schesnger, J . Brandriss, M. .,Walsh, E. E. 987 Pecion of miceagans dengue 2 virs encephalitis byimmunization with the dengue 2 virsnon-structura glycoproten N . Jen. Vrl. 6885357

43 Shege R W ed 980 he Tg-iruses Bilgy Sruure eplitin.

New York Academic 44. chlesnger, S. , chlesige, M. eds.1986. The Tgiride nd lir-ide. New York: Plenum

145 Schrader, A. P. esaway, E 1988.

Annu.R

ev.Microbiol.1990.44:649688


by

UniversityofMichigan-AnnArboron02/26/11.Forpersonalus

eonly.


39/40

VV NOM XPION 687

Tnlaio mapping with the flaviviruKunjin: ene order and anomalies ntranlaton of NS5 ru Re 932334

146 Smih W Wight 9 yn-

thesis f proens and glcopteins indengue e viruinfeted Ve adAede ptu cells J Gen. r66:5591

147 Speigh G, Coa G, ker M D,Wetaway G 1988 Gene mappingand positive denticaon of the non-sucul protein NS2A NS2B NSNS4B ad NS o he viviru unnand their eavage ie Gen. ro6923

148 Speight, G Westaway, G 1989Carboxytenal analsi of nne po-

teins sp(ced by he lavivis Kuninvden' ha only he intracellla oepoten is trunated Gen. r7022014

149 Sght Ww ostve entfcaton of S4A he lastf th hytha nntutual p-tein of vviruses roo 17099301

10 Srrairatna S Bhamrapravati N1 977 liaion of deng vru inAede aop uto Am. rop. Md. H. 26 1 191205

1 5 1 Srairaa S Bhamaapavati Nhalavadtana O 173 Denge virusinfeion f mie Mophogy and morphog f dngu y vu iscking mose nerones Infet. Immun. 8101728

152 Stehensn J Croks, A J LeeJ M 987 Th ynhei of imunogen polypeptide encoded bytickbo ncephalits vis J Genro 68130716

53 Sau G Strau H 1983Rpliatin tatge f h inglestranded RA viuses of eukaotes

urr r mmun 105198154 Strau G Stau J H 1985

Asemb o enveoped anma vsesIn ru truture n Aem, ed SJ Casen pp 20534 orola ValleyCA Jon and Batlett

15 Stas, G, Saus J H. 1986Suure nd replcaton of the alphavirs genome See ef 1 pp 3590

156 Staus H Sauss E. G 988volutn f RNA vu Ann. Re.Mroo. 4265783

157 Smosh Mo, c uke I,Moita uhra S e l 1987Compete nuceotide sequence of hapanese encephaltis vs genomeRN o 16147510

15 Svikin, Y V Lapstn V N, Lahkevch, V A Agol V I 1984 Dif-ferences between ranslation pdcts oftk nphlt vu RA n

cellfree stem fm Krebs2 clls andabi etiloyte nvolvemen ofmembrnes in he proessing of nascentprecror of aviviu sutral po-ens ro 133

159 Svkn Y . V gova Y Che-novskaa T V, Lapsn, V NLahkevich V A Agol V I 191Tlation o tickbo ncphlitivius favivius) genome n vtro Sntheis f wo ual polpepideroo 1 10234

160 Takegami T Washzu M sui

196 Nclotid ene a th 3 endof Japanee enephaliis vius genomeRNA rog 1524386

61 Takio Towatar T Katnuma N D C K. 198 Hlo-gy of amno acd sequences of at liverathepsin B and H wih tha of ppainPro. Natl Acd. Sci USA 80:366-

7016 Theier M Smith H H 1937 e of

low evr viru mdid b n trouvon f hu unztn xp. Md. 65787800

163 Tren D W Gran J A Monah T Manske, C , Cora M, o G 1989 Gneti vaiaton andievolution f dengue 2 vius nSotheast Asa 2

164 Tren D W Gran A Rosen L,Monah T 193 Gnetic vaitionang dngu f dffnt gogaphic origin ro 122714

165 Tren D W Grant J A VodamA V Monath T 191 Genetihetogneiy among Sint oui enpalitis viu ilte f dffent geographic ogn 143192

166 Trent D W Knne, R M JohnsonB J Vdam A V, Gant J A etal rtal nulode equne ofSt os encephaltis vius RNA:Sctral potens N , n2a andnsb roo 6293304

17 Trnt, D W, Nav C W 190Bhty and lition Se Rf108a, pp 1599

168 von Hene, G 94 ow sgnal seqenc inain cleavge speccityJ Mo. Bo. 1732451

19 Wahbg J M Boee W A M,

Garo 199 he heteodmeassociatin between the membrne pteins of Semki Foet viu changes tssenitivi to ow pH dung vusmaration r 6349197

Annu.R

ev.Microbiol.1990.44:649688.Downloadedfromwww.annualreviews.org

byUniversityofMichigan-AnnAr

boron02/26/11.

Forpersonalus

eonly.


40/40

88 HAM A

1 Wengler, G., Beato, M., Wengler G.199. In vitro anslaton of 42S vsspecc RNA om cells nfected wththe avvis West Nle vrus. Virology

951621 1 Wengler G, Castle E 196 Analyss

of suctral propertes whch possbyare charactersts for the 3 '-termnalsequences of the genome RNA of avvses Gen Vrol 6 1 1

1 Wenger, G Castle E, Ledner, Nowa T Wengler . 195 Sequence analyss of the membrane proteinV3 of th avvs West Nle s dof ts gene Virology 1

13 Wenger Wenger 191 Termnal sequences of the genome and re

atvor RNA of the favvsWest Nle vrs: Absence of poly(A) adpossble role n NA replcaton. Viology 1 13:555

14 Wengler G. Wengler G 199 Cellassoate Wet Nle avrs scovered th E+preM proten heterodimers which re destroyed ad reoganzed by proteolytc cleavage dungvrs l V 222

15 Wengler Wengler Nowk Wahn K 19 Analyss of the infuence of proteolytc cleavage on thestructral organaton of the surface ofthe West Nle avvs leads to thesolaton of a protease resstant E protenogomr from th vrl urc Voloy 16:119

16. Westaway E 19 Replcaton ofavvses. See Ref. 143 pp 5311

1 Wstwy 19 Favrsreplcato sraegy Adv Virus Res33459

1 Weaay E G Bnon M AGidmoich, Y. Honek M Igarash e al 19 FlvvaIntervirology 4:139

19 Wnler , en, F Kunz, C19 Stdes on the glycosylaton oflavvru E protns an he ole ofcarohydrte n antgenc stctre.Virology 159:23743

1 Wnler G Maxwell S. E Remmer C Stollr 199 Newly synthesed dng vrus nonscralproten NS s a soluble proten but beoms patally hydrophobc ad embneat aer dmezatonVoloy 1135

11 Wker G. Radoph eavesG R, yan T E. Stolla . 19Evdence that the maue form of thelavvrs nonscura roten NS 1 s a

dime. Viroogy 628912 World Health Organaton Scentc

roup. 195 Arthopod-Boe and odenBoe Viral Diease eneaWO

1. Wght P 192 Envelop proten ofthe favivirus Kunjn is apparently notglycosylated. J. Gen Virol. 59:29-38

14 Wght, P J Cauch M R Ng M L199 Dntn of the rboxy rmnof the tree glycoprotens speced byengue vs tp 2 VLo 111-

15 Wght P. J Warr H M. Westaway

E. 19 Prlmna haactezatonof glppt v m glcpens speced by the flavvrus Kunjn.Virology 19: 1

16 Wrght P. , Wa H M . , WestawayE . 191 Synthess f glycoprotensn cells nfected by the lavvs KunjnVirology 191

1 Yaegash, T akhara, . N, Page aa R . , Padanabhan R 986 Paial uotid sequene analyss of loned dengue rustp genome. Gene 6:56

1 amshchov F., Pletnev A G19. Nulotd sequece of thegenom regon encong he scuraptens n the NS1 prten of the tcboe encephalts vus. Nuleic Acide 165

19 Zhang . M., Hayes E P McCyT C Dubos D R Smers P Let a. 19 mmunaton of mce wthdengue structural protens d nonstructural proten NSI xprssed bybcovis ecombit indces esist-ance t ngu vu nphti Virol 6:33 1

19 Zha B Makow, E, BceWhteA Marko, L , Channo R. M . , etal. 196 Clonng ull length denge 4vral DNA seuences Analyss of genescodng for sctral prtens Virlogy155

191 hao B. Prnce " Qswood R cels K. Summers P . et a. 19Expresson of dengue vus suctuaprotes nd nonsucturl proten NS 1by a recombnt vacca Vl 6119

Annu.R

ev.Microbiol.1990.44:649688


by

UniversityofMichigan-AnnArboron02/26/11.Forpersonalus

eonly.

Documents

Chambers_et_al_-_Flavivirus_Genome_Organization_Expession_and_Replication_-_AnnuRevMicrobiol1990.pdf