MutationRank14-3-3sigma, BDS-2B99BAI1Noxap53AIP1PA26PCNAPiddPTGF-beta, FBS01RGCS100A2

wtR175H 1 - - - - - - - - - - -R248Q 2 - - - - - - - - - - -R273H 3 - - - - - - - - - - -R248W 4 - - - - - - - - - - -R273C 5 - - - - - - - - - - -R282W 6 - - - - - - - - - - -R249S 7 - - - - - - - - - - -G245S 8 - - - - - - - - - - -Y220C 9 - - - - - - - - - - -C176F 10 - - - - - - - - - - -V157F 11 - - - - - - - - - - -M237I 11 - - - - - - - - - - -E285K 13 - - -G245D 14 - - - - - - - - - - -H179R 15 - - - - - - - - - - -Y163C 16 - - - - - - - - - - -R273L 17 - - - - - - - - - - -Y234C 18 - - - - - - - -H179Y 19 - - - - - - - - - - -Y205C 20 - - - - - - - - - - -R248L 21 - - - - - - - - - - -V272M 23 - - - - - - -R158L 24 - - - - - - - - - - -C141Y 26 - - - - - - - - -P151S 27 - - - - - - - - - - -V173L 28 - - - - - - - - - - -R280T 30 - - - - - - - - - - -C238Y 32 - - - - - - - - - - -C242F 32 - - - - - - - - - - -P278S 35 - - - - - - - - - - -P278L 37 - - - - - - - - - - -C135Y 43 - - - - - - - - - - -H193R 45 - - - - - - - - - - -G244S 46 - - - - - - - - - - -R249M 50 - - - - - - - - - - -V173M 51 - - - - - - - - - - -K132R 56 - - - - - - - - - - -G244D 56 - - - - - - - - - - -M246V 61 - - - - - - - - - - -C135F 64 - - - - - - - - - - -K132N 68 - - - - - - - - - - -G279E 69 - - - - - - - - - - -M246I 74 - - - - - - - - - - -C238F 76 - - - - - - - - - - -H179L 81 - - - - - - - - - - -R249W 82 - - - - - - - - - - -P151H 87 - - - - - - - - - - -P177L 87 - - - - - - - - - - -N239S 87 - - - - - - - - - - -R273P 93 - - - - - - - - - - -C277Y 101 - - - - - - - - - - -R175L 106 -F270C 106 - - - - - - - - - - -R267W 119 - - - - - - - - - - -H168R 123 - - - - - - - - - - -D259Y 123 - - - - - - - - - - -H179N 128 - - - - - - - - - - -V143A 147 - - - - - - - - - - -L265P 151 - - - - - - - - - - -A276P 157 - - - - - - - - - - -R280S 157 - - - - - - - - - - -R158P 170 - - - - - - - - - - -P151R 187 - - - - - - - - - - -D281G 187 - - - - - - - - - - -G245R 202 - - - - - - - - - - -M246R 202 - - - - - - - - - - -L257Q 202 - - - - - - - - - - -C176R 219 - - - - - - - - - - -P278H 238 - - - - - - - - - - -L257P 256 - - - - - - - - - - -D281Y 277 - - - - - - - - - - -R283H 277 - - - - - - - - - -Y236D 340 - - - - - - - - - - -M237R 381 - - - - - -T256A 381 - - - - -S127P 417 - - - - - - - - - - -

loss of wt p53 function

Supp. Table I Dearth et al. 5

Mutation N C

wtR175H

R248Q

R273H

R248W

R273C

R282W

R249S

G245S

Y220C

C176F

V157F

M237I

E285K

G245D

H179R

Y163C

R273L

Y234C

H179Y

Y205C

R248L

V272M

R158L

C141Y

P151S

V173L

R280T

C238Y

P278L

C135Y

R249M

V173M

P151H

V143A

P151R

L257Q

Supp. Table II Dearth et al. 6

p53 DBS

MATa MAT

diploid

TRP1CEN

URA3

TRP1CEN

URA3

LEU2CEN

ADH1 p53

MATa

TRP1CEN

URA3

LEU2CEN

ADH1 p53

HIS3CEN

ADH1 p53mut

TRP1CEN

URA3

HIS3CEN

ADH1 p53mut

HIS3CEN

ADH1 p53mut

diploid

p53 mutant wt p53p53 DBS

A

Supp. Fig. 1 Dearth et al. 7

wt (eng)

R248QR273CR282WY220CV157FE285KY234CY205CR158LC141YP151SV272MP278LP278SH193Rwt (nat)

CS

R

cycl

in G

Fas

(AP

O-1

/CD

95)

GA

DD

45G

ML

KIL

LER

/DR

5M

DM

2m

dr1b

p21,

5’ s

itep2

1, 3

’ site

Type

IV C

olla

gena

se

wt (eng)R248QR273CR282WY220CV157FE285KY234CY205CR158LC141YP151SV272MP278LP278SH193Rwt (nat)

B

C

Supp. Fig. 1 Dearth et al. 8

wild

-type

R24

8QR

273C

R28

2WY

220C

V15

7FE

285K

Y23

4CY

205C

wild

-type

R15

8LC

141Y

P15

1SV

272M

P27

8LP

278S

H19

3R

D

Supp. Fig. 1 Dearth et al. 9

wt p53 NC

control NC

R175H NC

R248Q NC

R248W NC

R282W NC

R249S NC

wt p53 NC

C176F NC

M237I NC

E285K NC

H179R NC

G245D NC

H179Y NC

wt p53 NC

Y234C NC

R248L NC

R280T NC

P151S NC

C238Y NC

P278L NC

wt p53 NC

C135Y NC

P151H NC

V143A NC

C257Q NC

P151R NC

p

53

t

op

o II

t

ub

ulin

p

53

t

op

o II

t

ub

ulin

Supp. Fig. 2 Dearth et al. 10

2.51.5 2.0

Relative Activity

0 1.00.5

empty

wild-type

R282W

E285K

Y234C

V272M

R283H

M237R

2.51.5 2.00 1.00.5

empty

wild-type

R282W

E285K

Y234C

V272M

R283H

M237R

A

B

p21, 5’ site37˚C

Noxa37˚C

Supp. Fig. 3 Dearth et al. 11

A

B

2.01.5

Relative Activity

0 1.00.5

empty

wild-type

R282W

E285K

Y234C

V272M

p21, 5’ site30˚C

2.01.5

Relative Activity

0 1.00.5

empty

wild-type

R282W

E285K

Y234C

V272M

Noxa30˚C

0.2 0.6 1.0 1.4 1.61.2

Relative Activity

0 0.80.4

cyclin G

GADD45

GML

KILLER/DR5

MDM2

Pidd

C

30˚Cwild-typeR282W

wild-typeR282W

wild-typeR282W

wild-typeR282W

wild-typeR282W

wild-typeR282W

Supp. Fig. 4 Dearth et al. 12

Supp. Fig. 5

Relative Activity

mut

ant

wild

-typ

e

anti-

p53

R248Q

0.2 1.40.8 1.210

10

10

10

10

10

0

0.60.4

10

10

10

10

10

0

0

10

50

100

250

250

0

10

50

100

250

250

Noxa

PCNA

Dearth et al. 13

Class 1

Supp. Fig. 6

0.2 1.40.8 1.210 0.60.4

Class 3

mut

ant

wild

-typ

e

anti-

p53

anti-

p53

R282W

10

10

10

10

10

0

10

10

10

10

10

0

0

10

50

100

250

250

0

10

50

100

250

250

Noxa

PCNA

Relative Activity

Y234C

0.2 1.40.8 1.210 0.60.4

Relative Activity

0.2 1.40.8 1.210 0.60.4

Class 2

Relative Activity

mut

ant

wild

-typ

e

anti-

p53

R158L

10

10

10

10

10

0

10

10

10

10

10

0

0

10

50

100

250

250

0

10

50

100

250

250

Noxa

PCNA

R273C

Relative Activity an

ti-p5

30.2 0.8 1.210 0.60.4 1.4

Dearth et al. 14

Supp. Fig. 7GML

MDM2

Class 5

mut

ant

wild

-typ

e

10

10

10

10

10

0

10

10

10

10

10

0

0

10

50

100

250

250

0

10

50

100

250

250

Relative Activity

anti-

p53

Y220C

0.2 1.40.8 1.210 0.60.4

Relative Activity

anti-

p53

Y163C

0 0.2 1.40.8 1.210.60.4

Dearth et al. 15