Supplements to: A Nucleostemin-like GTPase Required for ... · A Nucleostemin-like GTPase Required for Apical and Floral Meristem Development in Arabidopsis X. Wang, D. K ... PKR

Supplements to: A Nucleostemin-like GTPase Required for Apical and Floral Meristem Development in

Arabidopsis X. Wang, D. K. Gingrich, Y. Deng and Z. Hong Table S1. Reduced floral organ numbers in nsn1 ap2 double mutants

Genotype

No. of flowers analyzed

No. of organ in whorl 1




WT 42 4 ± 0 4 ± 0 6 ± 0 2 ± 0

nsn1 42 2.2 ± 0.9 0.7 ± 1.2 2.4 ± 1.9 1.6 ± 0.7 ap2-1 42 4 ± 0 4 ± 0 5.9 ± 0 2 ± 0

nsn1 ap2-1 42 3.4 ± 0.9 1.6 ± 1.2 4.4 ± 1.7 2.0 ± 1.0 ap2-2 42 2.3 ± 0.7 0 ± 0 1.7 ± 1.2 1.9 ± 0.4

nsn1 ap2-2 42 1.5 ± 0.9 0 ± 0 0.1 ± 0.3 1.4 ± 0.8

1

AtNSN1Rice NSN1Amoeba NSN1Human NSMouse NSHuman GNL3LMouse GNL3LFly GNL3LAtNGP1Rice NGP1Human NGP1Mouse NGP1Fly NGP1

1 311 311 281 311 311 241 171 321 471 551 471 471 54

M V K R S - - - - - - - - - - - - - - - K K S K S K R V T L K Q - - - - - - - - - - - - K H K V L K K V K E H H K - - KM V K K S - - - - - - - - - - - - - - - K K S K S K R V T L R Q - - - - - - - - - - - - K H K V L R K V K E H H R - - KM G K Q T - - - - - - - - - - - - - - - - - - A S K R Q S L H H - - - - - - - - - - - - K H K V L K K V A E H H R - - KM K R P K - - - - - - - - - - - - - - - L K K A S K R M T C H K - - - - - - - - - - - - R Y K I Q K K V R E H H R - - KM K R P K - - - - - - - - - - - - - - - L K K A S K R M T C H K - - - - - - - - - - - - R Y K I Q K K V R E H H R - - K

M M K L R H K N K K - - - - - - - - - - - - P G E G S K G H K K I S W - - PM M K I R H K N K K - - - - - - - - - - - - P G K G S K - - - - - - - - - G

M A L K R - - - - - - - - - - - - - - L K T K K S K R L T G R L - - - - - - - - - - - - K H K I E K K V R D H N K - - KM V K K - - - - - E K K A N V S G K P K H S L D A N R A D G K K - - - - - - - K T T E T R S K S T V N R L K M Y K - T RM A K K K - - - - E R A V N V S G K P R H S L D V N R A N D K K G A G G G A G G G G G G R S A A T V R R L K M Y K - M RM V K P K - - - - Y K G R S T I N P S K A S T N P D R V Q G A G - - - - - - - - G Q N M R D R A T I R R L N M Y R - Q KM V K P K - - - - Y K G R S T I N R S A A S T N P D R V Q G A G - - - - - - - - G Q N M R D R G T I R R L N M Y R - Q KM P K V R S T T G K P R T Q G F N H S N H S M N P E R P K D G L K G - - - - - - V A H P R T K G T I K R L Q M Y R N F K


32 7832 7829 7432 7532 7525 6818 6133 7848 10456 11248 10748 10755 114

K A K D A K K L G L H R K P R V E K D P G - - - - - - I P N D W P F K E - Q E L K A L E V R R A R A L E E I - - - - - -K R K E A K K E G K S H R K K V E K D P G - - - - - - I P N E W P F K E - Q E L K A L E A R R A Q A L Q E L - - - - - -V K R F A K Q H P - - E L N K S R K D P G - - - - - - I P N L W P F K E D M L N K I E Q Q K Q K A S E E K K - - - - - -L R K E A K K Q G - - - H K K P R K D P G - - - - - - V P N S A P F K E - A L L R E A E L R K Q R L E E L K - - - - - -L R K E A K K R G - - - H K K P R K D P G - - - - - - V P N S A P F K E - A L L R E A E L R K Q Q L E E L K - - - - - -Y P Q P A K Q N G - - - K K A T S K V P S - - - - - - A P H F V H P N D - H A N R E A E L K K K W V E E M R - - - - - -C K K P A R Q N G - - - K K V T S R P S S - - - - - - A P Q I V H G N D - H A S R E A E L K K K R V E E M R - - - - - -E R R A A K K N P - - - - K K G S K K Q K - - - L I Q I P N I C P F K D - D I L K E V E E A K Q R Q E A E R - - - - - -P K R N A G G K I L S N E Y Q S K E L P N - - - S R I A P D R R W F G N T R V V N Q K E L E Y F R E E L Q T K M S S N YP L R D R G G K I L K H D L Q S K E L P N - - - T R I E P D R R W F G N T R V V N Q K E L E F F R E E L Q S R L S N N YE R R N S R G K I I K P L Q Y Q S T V A S G T V A R V E P N I K W F G N T R V I K Q S S L Q K F Q E E M D T V M K D P YE R R N S R G K V I K P L Q Y Q S T V A S G T V A R V E P N I K W F G N T R V I K Q A S L Q K F Q E E M D K V M K D P YA K R D R T G K I I C P A P F Q G R L A A G T M A R V E P T P K W F S N S R V I S Q T A L Q K F Q D E I G K A V K D P Y


79 10679 10675 10476 10376 10369 9762 9079 109105 162113 170108 167108 167115 172

- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - E Q K K E A R - K E R A K K R - K L G L V D D E D T K T E G- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - E L K K E A R - K E R A R K R - K L G L L E D E D I A N L A- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - N K K E K R R L E Q M A E A R R D I A S M A A D A K R R E S- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Q Q Q K L D R Q K E L E K K R - K L E T N - P D I K P S N V- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Q Q Q K L D R Q K E Q E R K R - K L E V S - P G D E Q S N V- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - E K Q Q A A R E Q E R Q K R R - T I E S Y C Q D V L R R Q E- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - E K Q Q V A R E Q E R Q R H R - T M E S Y C Q D V L K R Q Q- - - - - - - - - - - - - - - - - - - - - - - - - - - - - L A R R E A F K A E R E Q N K F K T L E S M V E D A D M R S TN V I L K E R K L P M S L L T D N K K - - Q S R V H L L D M E P F Q D A F G R K T K R K R P K L V A S D Y E A L V K K AN V I L K E R K L P L S L L Q D H Q K - - Q A R A H L L D T E P F E H A F G P K G K R K R P K L M A L D Y E S L L K K AK V V M K Q S K L P M S L L H D R I R P H N L K V H I L D T E S F E T T F G P K S Q R K R P N L F A S D M Q S L I E N AK V V M K Q S K L P M S L L H D R I Q P H N A K V H I L D T E S F E S T F G P K S Q R K R P N L F A S D M Q S L L E N AQ V I M K P S Q L P V T L L N E A S K - - Y K R V H L L D T E S F D S V F G P K K Q R K R V S L K V R D L E D L S K A A


107 143107 158105 151104 147104 14598 14191 134110 157163 222171 229168 223168 223173 228

E T I E D L P - - - - - - - - - - - - - - - - - - - K V V N V R D N S E - - - - R A F Y K E L V K V I E L S D V I L E VS A A S A Q G S E F - - - - A E K D A A K E N A S L D V V K S Q D H S E - - - - R A F Y K E L V K V I E A S D V I L E VE F Q E R Q Q L K - - - - - - - - - - - - Q Q Q K Q Q G K F E K E G K D - S S L K Q F Y R E V K K V I E A G D V I L Q VE P M E K E F G L - - - - - - - - - - - - C K T E N K A K S G K Q N S K - - - - K L Y C Q E L K K V I E A S D V V L E VE T R E E S D - - - - - - - - - - - - - - E P K R K K A K A G K Q N P K - - - - K L H C Q E L K K V I E A S D I V L E VE F E H K E E V L - - - - - - - - - - - - Q E L N M F P Q L D D E A T R - - - - K A Y Y K E F R K V V E Y S D V I L E VE F E Q K E E V L - - - - - - - - - - - - Q E L N M F P Q L D D E A T R - - - - K A Y Y K E F R K V V E Y S D V I L E VV H G I M H E N D - - - - - - - - - - - - A Q D Q D E K K Y K N A V T K E Q S L K Q Y F K E F R K V I E N A D V V L E VA E S Q D A F E E K N G A G P S G E G G E E E D G F R D L V R H T M F E K G Q S K R I W G E L Y K V I D S S D V I V Q VD D S Q G A F E D K H A T A K L L K E - E E E D G L R D L V R H T M F E K G Q S K R I W G E L Y K V I D S S D V V V Q VE M S T E S Y D Q - - - - G K D R D L V T E D T G V R N E A Q E E I Y K K G Q S K R I W G E L Y K V I D S S D V V V Q VE M S T E S Y D Q - - - - G K D R D L V M E D T G V R N E A Q E E I Y K K G Q S K R I W G E L Y K V I D S S D V V V Q VG E Q A D K Y D S - - - - T K D I D L I R E D T G E K K A V R D W V F G A G G S R R I W N E L H K V V D A S D V L L Q V


144 203159 218152 211148 206146 204142 201135 194158 217223 282230 289224 283224 283229 288

L D A R D P L G T R C T D M E R M V M Q A G P N K H L V L L L N K I D L V P R E A A E K W L M Y L R E E F P A V A F K CL D A R D P L G T R C I D M E K M V R K A D P S K R I V L L L N K I D L V P K E S V E K W L T Y L R E E M P T V A F K CL D A R D P M G C R C L E I E K M I L E R Y T N K K I V L I L N K I D L V P R E N V L M W L K Y L R N F Y P T L A F K CL D A R D P L G C R C P Q V E E A I V Q S - G Q K K L V L I L N K S D L V P K E N L E S W L N Y L K K E L P T V V F R AL D A R D P L G C R C P Q I E E A V I Q S - G S K K L I L V L N K S D L V P K E N L E N W L N Y L N K E L P T V V F K AL D A R D P L G C R C F Q M E E A V L R A Q G N K K L V L V L N K I D L V P K E V V E K W L D Y L R N E L P T V A F K AL D A R D P L G C R C F Q M E E T V L R A E G N K K L V L V L N K I D L V P K E I V E K W L E Y L L N E L P T V A F K AV D A R D P L G T R C N E V E R A V R G A P G N K R L V L V L N K A D L V P R E N L N N W I K Y F R R S G P V T A F K AI D A R D P Q G T R C H H L E K T L K E H H K H K H M I L L L N K C D L V P A W A T K G W L R V L S K E Y P T L A F H AL D A R D P M G T R C Y H L E K H L K E N A K H K H L V F L L N K C D L V P A W A T K G W L R T L S K D Y P T L A F H AL D A R D P M G T R S P H I E T Y L K K E K P W K H L I F V L N K C D L V P T W A T K R W V A V L S Q D Y P T L A F H AL D A R D P M G T R S P H I E A Y L K K E K P W K H L I F V L N K C D L V P T W A T K R W V A V L S Q D Y P T L A F H AL D A R D P M G T R S K Y I E E F L R K E K P H K H L F F I L N K V D L V P V W V T Q R W V A I L S A E Y P T I A F H A


204 260219 275212 270207 261205 256202 259195 253218 274283 316290 323284 317284 317289 322

S T Q E - Q R S N L G W K S S K A S K P S - - N M L Q T S D C L G A D T L I K L L K N Y S R S H E L K K S I T V G I I GN T Q E - Q R T K L G W K S S K I D K S S - - N I P Q S S D C L G A E N L I K L L K N Y S R S H E L K L A I T V G I V GS T Q Q - Q K R N L G Q Q G G I Q P E L A S N D M L N S T E S L G A E Q L L Q L L K N Y S R S L N I K T S V T V G I I GS T K P - - - K D K G K I T K R V K A K K N A A P F R S E V C F G K E G L W K L L G G F Q E T - - C S K A I R V G V I GS T N L - - - K N R K - - T F K I K K K K - V V P F Q S K I C C G K E A L W K L L G D F Q Q S - - C G K D I Q V G V I GS T Q H - Q V K N L N R C S V P V D Q A S - E S L L K S K A C F G A E N L M R V L G N Y C R L G E V R T H I R V G V V GS T Q H H Q V K N L T R C K V P V D Q A S - E S L L K S R A C F G A E N L M R V L G N Y C R L G E V R G H I R V G V V GS T Q D Q A N R L G R R - - - K L R E M K T E K A M Q G S V C I G A E L L M S M L G N Y C R N K G I K T S I R V G V V GS V N K - - - - - - - - - - - - - - - - - - - - - - - - - - S F G K G S L L S V L R Q F A R L K S D K Q A I S V G F V GS I N S - - - - - - - - - - - - - - - - - - - - - - - - - - S F G K G S L L S V L R Q F A R L K S D K Q A I S V G F V GS L T N - - - - - - - - - - - - - - - - - - - - - - - - - - P F G K G A F I Q L L R Q F G K L H T D K K Q I S V G F I GS L T N - - - - - - - - - - - - - - - - - - - - - - - - - - P F G K G A F I Q L L R Q F G K L H T D K K Q I S V G F I GS L Q H - - - - - - - - - - - - - - - - - - - - - - - - - - P F G K G A L I N L F R Q L G K L H L D K K Q I S V G F I G


261 320276 335271 329262 320257 315260 317254 311275 333317 375324 382318 376318 376323 381

L P N V G K S S L I N S L K R A H V V N V G A T P G L T R S L Q E V H L D K N V K L L D C P G V V M L K S S G N D A S IL P N V G K S S L I N S L K R S R V V N V G S T P G V T R S M Q E V Q L D K K V K L L D C P G V V M L K S S N S G V S VY P N V G K S S L I N S L K R T R S V G V G A T P G F T K F A Q E V H L D K N V K L L D S P G I V P I K G - N V D E N IF P N V G K S S I I N S L K Q E Q M C N V G V S M G L T R S M Q V V P L D K Q I T I I D S P S F I V S P L - N S S S A LF P N V G K S S V I N S L K Q E W I C N V G I S M G L T R S M Q I V P L D K Q I T I I D S P C L I I S P C - N S P T A LL P N V G K S S L I N S L K R S R A C S V G A V P G I T K F M Q E V Y L D K F I R L L D A P G I V P G P - - N S E V G TL P N V G K S S L I N S L K R S R A C S V G A V P G V T K F M Q E V Y L D K F I R L L D A P G I V P G P - - N S E V G TI P N V G K S S I I N S L T R G R S C M V G S T P G V T K S M Q E V E L D S K I K L I D C P G I V F T S G - G E N S H AY P N V G K S S V I N T L R T K N V C K V A P I P G E T K V W Q Y I T L T K R I F L I D C P G V V Y Q S R - D T E T D IY P N V G K S S V I N T L R S K S V C K V A P I P G E T K V W Q Y I T L T K R I F L I D C P G V V Y Q N N - D S E T D IY P N V G K S S V I N T L R S K K V C N V A P I A G E T K V W Q Y I T L M R R I F L I D C P G V V Y P S E - D S E T D IY P N V G K S S V I N T L R S K K V C N V A P I A G E T K V W Q Y I T L M R R I F L I D C P G V V Y P S E - D S E T D IY P N V G K S S V I N A L R S K K V C K V A P I A G E T K V W Q Y I T L M K R I F L I D C P G V V Y P T A - E T D T E K

G5 G4

G1 G2 G3

2


321 380336 395330 389321 379316 374318 377312 371334 393376 435383 442377 436377 436382 441

A L R N C K R I E K L D D P V S P V K E I L K L C P K D M L V T L Y K I P S F E A V D D F L Y K V A T V R G K L K K G GA L R N C K R V E K M E D P I S P V K E I L D L C P H E K L L S L Y R V P T F T S V D D F L Q K V A T L R G K L K K G GI L R N V V K L E K V L D P I A P V D A I L S R C S Q K Q I L D I Y E I A Q Y Q S T T D F L T Q V A A K R K K I V K G GA L R S P A S I E V V K - P M E A A S A I L S Q A D A R Q V V L K Y T V P G Y R N S L E F F T M L A Q R R G M H Q K G GA L R S P A S I E E L R - P L E A A S A I L S Q A D N E Q V V L K Y T V P E Y K D S L H F F T K L A Q R R G L H Q K G GI L R N C V H V Q K L A D P V T P V E T I L Q R C N L E E I S N Y Y G V S G F Q T T E H F L T A V A H R L G K K K K G GI L R N C I H V Q K L A D P V T P V E T I L Q R C N L E E I S S Y Y G V S G F Q T T E H F L T A V A H R L G K K K K G GV L K N A Q R V G D V K D P F T I A E S V L K R A S K E Y F C T M Y D I T N Y D T F E E F F A K K A A R M G K F L K K GV L K G V V R V T N L E D A S E H I G E V L R R V K K E H L Q R A Y K I K D W E D D H D F L L Q L C K S S G K L L K G GV L K G V V R V T N L A D A S E H I G E V L R R V K K E H L K R A Y K I E D W V D D N D F L V Q L S K T T G K L L R G GV L K G V V Q V E K I K S P E D H I G A V L E R A K P E Y I S K T Y K I D S W E N A E D F L E K L A F R T G K L L K G GV L K G V V Q V E K I K A P Q D H I G A V L E R A K P E Y I S K T Y K I E S W E N A E D F L E K L A L R T G K L L K G GV L K G V V R V E L V T N P E D Y V D T V L Q R V R K E Y I A K N Y K I D D W T S P K H F L E Q L A Q K S G K L L K G G


381 421396 436390 431380 421375 416378 419372 413394 436436 472443 480437 496437 496442 481

L V D I D A A A R I V L H D W N E G K I P Y Y T M P P K - - - - - - - - - - - - - - - - - - - R D Q G G H A E S K I V TI V D V E A A A R I V L H D W N E G K I P Y F T V P P K - - - - - - - - - - - - - - - - - - - R D A V D D S D A V I I SI A D L R S T A I S V I R D W T G G K I P F Y T Q P P K - - - - - - - - - - - - - - - - - - V L V K S T L L S Q F S D EI P N V E G A A K L L W S E W T G A S L A Y Y C H P P T S - - - - - - - - - - - - - - - - - - W T P P P Y F N E S I V VS P N V E S A A K L V W S E W T G A S L G Y Y C H P P A S - - - - - - - - - - - - - - - - - - W N H S L H F N E N I A AL Y S Q E Q A A K A V L A D W V S G K I S F Y I P P P A T - - - - - - - - - - - - - - - - - - H T L P T H L S A E I V KV Y S Q E Q A A K A V L A D W V S G K I S F Y T L P P P T - - - - - - - - - - - - - - - - - - H T L P T H L S A E I V KV P D V V A A A R S V L N D W N T G K I K Y C T Q P P E V Q - - - - - - - - - - - - - - - - - E G Q S V H I S A S I V HE P D L M T G A K M I L H D W Q R G R I P F F V P P P - - - - - - - - - - - - - - - - - - - - K - L D N V A S E - - S EE P D L T T T A K M V L H D W Q R G K I P F F V P P P - - - - - - - - - - - - - - - - - - - - Q Q G E D S P S E - - T AE P D L Q T V G K M V L N D W Q R G R I P F F V K P P N A E P L V A P Q L L P S S S L E V V P E A A Q N N P G E E V T EE P D M L T V S K M V L N D W Q R G R I P F F V K P P N A E I P T D S Q L P P S S P L E V P T E T T Q N N P E E E T T EE P D V T V T S R M V L N D W Q R G K L P F Y V P P E - - - - - - - - - - - - - - - - - - - - G F A T P K S Q E D K V S


422 459437 474432 471422 456417 451420 466414 460437 472473 507481 514497 546497 546482 523

E L A K D F N - - I D E V Y S G E S - - - - - - - - - S F I G S L K T - - - - - - - - - - - V N E F N P V I I P S N G PE T G K E F N - - I D E I Y K A E S - - - - - - - - - S Y I G G L K S - - - - - - - - - - - L E E F R H I E I P S N A PL N I D Q S D - - L I S T V S N Q N - - - - - - - - - T S F A S L K Y L E P K - - - - - - - Q Q E L D N S L F D D D - -D M K S G F N - - L E E L E K N N A - - - - - - - - - Q S I R A I K G - - - - - - - - - - - P H L A N S I L F Q S - - -V M K K G F N - - L E E L E K N N A - - - - - - - - - H S I Q V L K G - - - - - - - - - - - P H L T N R I L F R S - - -E M T E V F D - - I E D T E Q A N E - - - - - - - - - D T M E C L A T G E S D E L L G D T D P L E M E I K L L H S P - -E M T E V F D - - I E D T E H A N E - - - - - - - - - D T M E C L A V G E S D E L L G D M D P Q E M E V R W L H S P - -S E A R E F D - - V E N F E S M E T - - - - - - - - - E I L E H C A V - - - - - - - - - - - - K T D D I M E I T S T G -V I V P G I D - - - K E A I A D N S - - - - - - - - - Q A A A A L K A I - - - - - - - - - - A G I M S T Q Q Q K D - - -E P V E K S D - - - E E G V S S D - - - - - - - - - - R T A A A M K A I - - - - - - - - - - A G I I S S Q Q Q M N - - -T A G E G S E S I I K E E T E E N S H C D A N T E M Q Q I L T R V R Q N - - - - - - - - - - F G K I N V V P Q F S G D DT E V E R S D S I T E K E P E G D C S Q D R N S E M Q Q I L A R V R Q N - - - - - - - - - - F G K I N V G P Q F S A D DE E A A G D D A N S D A K S E A P T F V S - - - - - - E S V K R A R E F - - - - - - - - - - K Q I Q D F R K I R V G - -


460 518475 526472 524457 510452 491467 519461 514473 524508 557515 527547 599547 599524 575

- L N F D E T M I E D E S K T Q T E E E A E H E S D D D E S M G G E E E E E A G K T K E K S E T G R Q N V K L Y A A E S- P Q I D E E M L E D - G K K Q N E P A Q E N - - - H D E S M S D A N E R E G A K T A S A S - - - T Q N D K L Y T A E G- D E E D E E E M E D - - E D E D D D E E E E - - - - E E M E D E E D E E D E E D E E D E D E M E E D S D S E A L L K Y- S G L T N G I I E E - - K D I H E E L P K R - - - K E R K Q E E R E D D K D S D Q E T V D E E V D E N S S G M F A A E- S G L T N G I L D E - - K D I V E E S P - - - - - - - - - - S Q T E D Q Q D A D D - - - - - - - Q E N G S G E R N A E- M T K I A D A I E N - - K T T V Y K I G D L - - - T G Y C T N P N R H Q M G W A K R N V D H R - P K S N S M V D V C S- L V K I A D A I E N - - R S T V Y K I G N L - - - T G Y C T K P N R N Q M G W P K R N V D H H C P Q N N R V V E V S S- P L E I R Q P R E E - - A E P A D K I T A S - - - - - L V I D E K E K P A K G R K R K L D E E K E K V D P S L L L E E- V P V Q R D F Y D E - - K D L K D - - - D K - - - - K A K E S T E T D A E N G T D A E E D E D A V S E D G V E S D S D- V P C Q K E F G - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - V T N E D - - - - - - -L V P V E V S D L E E - - E L E S F S D E E E - - - - E E Q E Q Q R D D A E E S S S - E P E E E N V G N D T K A V I K AL V P V E M S D L E D - - L E S S G - E E E E - - - - Q E Q E Q P G E D A E E E R S P D T Q E E P V G N D T K A V L R A- L E Y E Q Q D V K E - - L D H I D L E L L E - - - - Q - Q K A E R A A R K K A R Q L N L D D E E E S S D G A S D F Y S


519 559527 567525 580511 544492 519520 556515 551525 561558 576528 535600 653600 652576 630

M L N T K K Q K A E K K K R K K A K K A G A D E E D L - - M D G D Y D F K V D Y R K N - - - - - - - - - - - - - - - - -I L D P R K R K A E K K - R R K A N K F S V L T D - - - - M D A D Y D F K V D Y Q M K D A L - - - - - - - - - - - - - -Q Q Q K Q V V K A T A K Q V A Q I K S K S Q N L K D E N D Q F N P Q S N K Q L K K Q N K K L K K K F G I V S T S - - - -E T - - - - G E A L L R R L Q Q V N S L Q G L L S - - - - W I K S L K R M M L M T S - - - - - - - - - - - - - - - - - -I S - - - - D V A P V E E T R E L S P E Q - - - - - - - - - - S T A G K P S D G S S - - - - - - - - - - - - - - - - - -V D - - - - R R S V L Q R I M E T D P L Q Q G Q A - - - - L A S A L K N K K K M Q K R A D - - - - - - - - - - - - - - -V D - - - - R R P M L Q R I L E T D P L Q Q G Q A - - - - L A S A L K N K K K L Q K R S D - - - - - - - - - - - - - - -N Q - - - S L N K G I K Q M Q K L K K K Q N V R N E K - - K I S K I T D V L D S F S - - - - - - - - - - - - - - - - - -A D - - - E D A V S E N D E E D - E S D S A E- - - - - S E V A E Q S EL D - - - E K I A K Y Q K F L D - K A K A K K F S A V - - R I S K G L S E K I F A K P E E Q R K T L E E D V D D R A P SL D - - - E K I A K Y Q R F L N - K A K A K K F S A V - - R I S K D L S E K V F A K Y K E E - K K T S A E V S D A A P TE D - - - E Y N D D L Q R V V H R K A K T K K I Q Q L - - A I T S S G K F R V A K V Q P G D S D D M G N S S D D A A G P


560 582568 591581 615545 560520 538557 582552 577562 581577 576536 535654 713653 710631 674

- - - - - - - - - - - - - - - - - - - - - - - - K D G E D E E F Q I D A K I P M A G L L P E E- - - - - - - - - - - - - - - - - - - - A E E G N D G G D E - - E P K E A D P M T G V D D A- - - - - - - - - - - - - - - - - - - - S K Q V Q D D D D E - - D D D D E Q E E I D Q D D E D D E D I D E D E A F- - - - - - - - - - - - - - - - - - - - - - - V Q I M C N R - - - T M A F Y D F F F- - - - - - - - - - - - - - - - - - - - - - - A L D R A S Q - - - E D E T Y D F T T D Y I- - - - - - - - - - - - - - - - - - - - - K I A S K L S D S - - - M M S A L D L S G N A D D G V G D- - - - - - - - - - - - - - - - - - - - - K I A T K L S D S - - - M M S M L D L S G N S D D C A G D- - - - - - - - - - - - - - - - - - - - - - - L G P S S S K - - - - A E K Y D F D E D Y V I E

K K G K K R K A Q R E E E Q E H S N K A P R A L T S K E R R R A V R Q Q R P K K V G V R Y Y E T H N V K N R N R N K K KK K A R K W D A Q M E E E P - - S N K T Q R M L T C K E R R R A A R Q Q Q S K K V G V R Y Y E T H N V K N R N R N K K KS S A K - - - - - - - - - - - - - - - - S P R L T A K Q K R A L E R N Q K R K K I G S N F Y E M T N V K N R N R N K K R


583 582592 591616 615561 560539 538583 582578 577582 581577 576536 535714 731711 728675 675

T N D S E G Q K H K R K K F R Q K QT S D S E G Q K H R R N K F R Q K QD

Suppl Figure S1. Amino acid sequence alignment of NSN1 homologs.

Dark shades indicate identical residues appearing in at least 7 peptide sequences, and light shades indicate sim-ilar residues appearing in at least 7 peptides. GTP binding sundomains (G1 -G5) are marked with green lines.

3

Suppl. Figure S2. Growth and reproduction phenotypes of nsn1-2 mutant plants. (A-B) 20-day-old plants of the WT control (A) and nsn1-2 (B). (C) Siliques of nsn1-2 containedaborted ovaries and produced few seeds, as compared to the wild type (WT). Arrow-heads indicate the aborted ovaries in nsn1-2 homozygote siliques. (D) Indeterminateinflorescence of a 35-day-old wild type plant. (E) Terminating inflorescence withcarpelloid flowers of a 45-day-old nsn1-2 mutant plant. (F) An enlarged defectiveflower from (E). Scale bars, 5 mm in (A-B), 2 mm in (C-F).

(A) (B) (C)

(D) (E) (F)WT nsn1-2 WT nsn1-2

WT nsn1-2 nsn1-2

4

WT nsn1/+

1050

520

+ + + + ++ + + ++ + + +

nsn1

Lb1LPRP

(B)

(A)

nnnnssssnnnn1111----1111Nsn1-LP

Lb1 Lb1

Nsn1-R

1050 bp770 520

ATG

nnnnssssnnnn1111----2222

5' 3'

Nsn1-F

N857-R1Nsn1-RP

2.0

1.00.6

0.10.2

* *

WT nsn1 WT nsn1 WT nsn1

Nsn1-F/ Nsn1-R

Nsn1-LP/ Nsn1-RP

Nsn1-F/ N857-R1

NSN1

NSN1-3'

NSN1-5'primer

(C)

NSN1

ACTIN

Rl Ia St Fl Si(D)

(G)(F)

236

23

7

IM(E) (H)

GFP NSN1OXWT nsn1-1

IM

62

3

6

Suppl. Figure S3. Expression of NSN1 mRNA in wild type Arabidopsis, nsn1-1 mutant and NSN1OX plants. (A)Positions of three pairs of primers used to detect NSN1 mRNA transcript in nsn1-1 mutant plants. PrimersNsn1-F and Nsn1-R correspond to the cDNA sequences in the start codon and stop codon regions, and pro-duce a PCR product of 1,750 bp using genomic DNA of WT plants. Primers Nsn1-LP and Nsn1-RP flank theT-DNA insertion site and produced a 1050-bp PCR product in WT. Nsn1-F and N857-R1 amplify a 256-bpgenomic DNA product or a 133-bp cDNA band in WT plants. Filled boxes are coding exons of NSN1 geneand open boxes are non-coding exons. Lines represent introns of NSN1. (B) Genomic DNA isolated fromindividual plants was used for PCR amplification using primers as indicated in (A). (C) RT-PCR results usingthree pairs of primers. Note that the 5'-mRNA coding sequence, the mRNA region flanking the T-DNA inser-tion site, and the full-length NSN1 mRNA were undetectable in nsn1-1 mutant, suggesting that it is a null-mutation. Primer dimers are indicated by asterisks. (D) Tissue-specific expression of NSN1 using RT-PCR.RT-PCR was done using total RNAs from different tissues, including rosette leaf (Rl), inflorescence apex (Ia),stem (St), flower (Fl), and silique (Si). The expression of Actin gene served as an internal template control.(E-H) Expression levels of NSN1 mRNA in the inflorescence meristem of wildtype (E), nsn1-1 (F), andtransgenic plants expressing 35Spro:GFP (a control, G) and 35Spro:NSN1-GFP (NSN1OX, H). In the controlsof wild type and GFP-expressing plants, the NSN1 mRNA signals could be detected in the IM and floral pri-mordia at stages 2-6. In NSN1OX transgenic plants, the NSN1 mRNA signal was enhanced, and could bedetected in the IM and floral primordia of various stages. The NSN1 mRNA could also be detected in thepedicles of developing flowers. Scale bars, 100 μm in (E-H).

5

seedlingleaf

inflorescencestemflowercarpelovarypetalsepalpollensiliqueseed

shoot apexroot

0 1000 2000 3000Relative expression level of NSN1 (Log2)

Suppl. Figure S4. Expression profile of NSN1 mRNA in different tissues ofwild type Arabidopsis plants. The expression level of At3g07050 (NSN1)is presented in relative values in log2. Data were retrieved from the publicmicroarray data set (https://www.genevestigator.ethz.ch).

6

Suppl. Figure S5. Non-terminal carpelloid flowers of nsn1-1. Carpelloid flowers wereproduced at non-terminal positions of inflorescences of nsn1-1 plants. Shown werea carpelloid flower at position 38 (A) and position 35 (B) of a 40 day-old nsn1-1plant, which developed a terminal carpelloid flower at the position of flower num-ber 41.

(B)(A)

nsn1nsn1

7

0

5

10

15 *

WT nsn10

20

40

60 *

WT nsn1

(D)(C)

(A) (B)

1

1

2

2 3

3

4

4

5

5

Suppl. Figure S6. Reduced size of spical domes in nsn1. A-B, Scan electron microscope (SEM)images of the wild type (A) and nsn1-1 (B) shoot apical meristems (SAM). Floral primordiaare successively numbered from 1 to 5. The lines indicate the distance along which meristemsizes and meristem cell numbers were measured. C-D, Quantification of the apical domes. Thelength of L1 layer epidermal cells (C) and the number of epidermal cells along the line (D) ofeight plants for each genotype were analyzed. A star (*) indicates a statistically significant dif-ference between nsn1 and the wide type (T-test proceeded by Origin 6.0, P<0.01).

WT nsn1

8

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Supplements to: A Nucleostemin-like GTPase Required for ... · A Nucleostemin-like GTPase Required for Apical and Floral Meristem Development in Arabidopsis X. Wang, D. K ... PKR