TABLE 1. Examples of p53-interacting Proteins p53 domain
involved in Selected
p53-interacting protein Method interaction references
A ctivating Tr anscription F actor 3 (ATF3) GST pull-down,co-IP FL 1 A
+ taxia- T elangiectasia- M utated (ATM) GST pull-down FL 2 Bcl-XL co-
+IP,GST pull-down DNA-binding domain 3
Collaborator of ARF (CARF) co-IP FL 4
Ch eckpoint k inase 2 (Chk2) GST pull-down,co-IP amino terminal 5,6 Co
+ nstitutively P hoto-morphogenic 1 (COP1) IP-tagged protein,silver st
+aining 7 + mass spectrometry G lycogen S ynthase K inase-3 
 (GSK3 
 
+), IP carboxyl terminal 8 H erpesvirus- A ssociated U biquitin- S pec
+ific mass spectrometry of affinity- P rotease (HAUSP) purified p53-as
+sociated factors FL 9
H uman ortholog, S ilent i nformation r egulator (hSir2) co-IP,IF carb
+oxyl terminal and central 10 c- J un- N -terminal k inase (JNK) co-IP
+ amino terminal 11 M uscle S egment homeodomain family,vertebrate co-
+IP FL 12 homolog 1 (Msx1)
N ADPH: q uinone o xidoreductase 1 (NQO1) gel filtration,co-IP FL 13,1
+4 P oly( A DP- R ibose) P olymerase-1 (PARP-1) IP central and carboxy
+-terminal 15 fragments P rotein I nhibitor of A ctivated S TAT (PIAS)
+ Y2H,co-IP FL 16,17 p 53- i nduced protein with a R ING H2 domain (Pi
+rh2) co-IP DNA-binding domain 18 P ro m yelocytic l eukemia protein (
+PML) GST pull-down and IVT DNA-binding domain 19 P300/CBP co-IP,ChIP 
+amino terminal 20
P olo- l ike k inase 1 (Plk1) co-IP DNA-binding domain 21 Re dox f act
+or 1/ AP - E ndonuclease 1 (Ref-1/APE1) far-western and IP-western FL
+ 22 assays in vitro S caffold/ M atrix A ttachment R egion binding co
+-IP FL 23 protein-1 (SMAR1)
S100B co-IP FL 24
T BP- A ssociated F actor 1 (TAF1) co-IP carboxyl terminal 25 Tr ansfo
+rmation/t r anscription domain a ssociated co-IP,GST pull-down carboo
+xyl terminal 26 p rotein (TRRAP),hGcn5,TAF II 30 W e rn ers�sy
+ndrome protein (WRN) co-IP FL 27
14-3-3 co-IP,GST pull-down carboxyl terminal 28 The proteins listed in
+ this table represent some (not all) of the factors identified as p53
+ interactors since 1998. The significa nt majority of these inter- ac
+tions were identified and characterized by targeted co-IP or by other
+ biochemical means; in contrast, two-hybrid and other gene tic approa
+ches have been less useful for identification of p53 interactors.IP,i
+mmunopr ecipitation;Y2H,yeast two-hybrid; IVT,in vitro translation; C
+hIP ,chromatin IP; FL, full length;IF,immunofluorescence. R EFERENCES
(1) Yan C,Lu D,Hai T,and Boyd DD.2005.Activating transcription factor
3,a stress sensor,activates p53 by blocking its ubiquitination. EMBO J
+. 24: 2425�2435. (2) Khanna K.K., Keating K.E., Kozlov S., Sco
+tt S., Gatei M., Hobson K.,
Taya Y., Gabrielli B., Chan D., Lees-Miller S.P., and Lavin M.F. 1998.
ATM associates with and phosphorylates p53: Mapping the region of
interaction. Nat.Genet. 20: 398�400. (3) Mihara M.,Erster S.,Za
+ika A.,Petrenko O.,Chittenden T.,Pancoska P.,
and Moll U.M. 2003. p53 has a direct apoptogenic role at the mito-
chondria. Mol.Cell 11: 577�590. (4) Hasan M.K.,Yaguchi T.,Minod
+a Y.,Hirano T.,Taira K.,Wadhwa R.,and
Kaul S.C . 2004.Alternative reading frame protein (ARF)-independent fu
+nction of CARF (collaborator of ARF) involves its interactions with
p53:Evidence for a novel p53-activation pathway and its negative feed-
back control. Biochem.J. 380: 605�610. (5) Shieh S.Y., Ahn J., 
+Tamai K., Taya Y., and Proves C. 2000. The human
homologs ofcheckpoint kinases C hk1 and Cds1 (Chk2) phosphorylate p53 
+at multiple DNA damage-inducible sites. Genes Dev . 14: 289�30
+0. (6) Berger M.,Stahl N.,Del Sal G.,and Haupt Y.2005.Mutations in pr
+oline
82 of p53 impair its activation by Pin1 and Chk2 in response to DNA da
+mage. Mol.Cell.Biol. 25: 5380�5388. (7) Dornan D., Wertz I., S
+himizu H., Arnott D., Frantz G.D., Dowd P.,
O�Rourke K., Koeppen H., and Dixit V.M. 2004 . The ubiquitin li
+gase COP1 is a critical negative regulator of p53. Nature 429: 86&#65
+533;92. (8) Watcharasit P.,Bijur G.N.,Song L.,Zhu J.,Chen X.,and Jope
+ R.S.2003.
Glycogen synthase kinase-3beta (GSK3beta) binds to and promotes the
actions of p53. J.Biol.Chem. 278: 48872�48879. (9) Li M.,Chen D
+.,Shiloh A.,Luo J.,Nikolaev A.Y.,Qin J.,and Gu W.2002.
Deubiquitination of p53 by HAUSP is an important pathway for p53
stabilization. Nature 416: 648-653. ( 10) Vaziri H.,Dessain S.K.,Ng Ea
+ton E.,Imai S.I.,Frye R.A.,Pandita T.K.,
Guarente L., and Weinberg R.A. 2001. hSir2(SIRT1) functions as an
NAD-dependent p53 deacetylase. Cell 107: 149�159. (11) Fuchs S.
+Y.,Adler V.,Buschmann T.,Yin Z.,Wu X.,Jones S.N.,and Ronai
Z.1998.JNK targets p53 ubiquitination and degradation in nonstressed
cells. Genes Dev. 12: 2658�2663. (12) Park K.,Kim K.,Rho S.B.,C
+hoi K.,Kim D.,Oh S.H.,Park J.,Lee S.H.,
and Lee J.H.2005.Homeobox Msx1 interacts with p53 tumor suppres-
sor and inhibits tumor growth by inducing apoptosis. Cancer Res. 65: 7
+49�757.
(13) Anwar A., Dehn D., Siegel D., Kepa J.K., Tang L.J., Pietenpol J.A
+., and
Ross D.2003.Interaction ofhuman NAD(P)H:quinone oxidoreductase
1 (NQO1) with the tumor suppressor protein p53 in cells and cell-free
systems. J.Biol.Chem. 278: 10368�10373. (14) Asher G., Tsvetkov
+ P., Kahana C., and Shaul Y. 2005. A mechanism of
ubiquitin-independent proteasomal degr adation ofthe tumor suppres- so
+rs p53 and p73. Genes Dev. 19: 316�321. (15) Wesierska-Gadek J
+., Wojciechowski J., and Schmid G. 2003. Phos-
phorylation regulates the interaction and complex formation between
wt p53 protein and PARP-1. J.Cell.Biochem. 89: 1260�1284. (16) 
+Gallagher W.M., Argentini M., Sierra V., Bracco L., Debussche L., and
Conseiller E.1999.MBP1:A novel mutant p53-specific protein partner
with oncogenic properties. Oncogene 18: 3608�3616. (17) Okubo S
+.,Hara F.,Tsuchida Y.,Shimotakahara S.,Suzuki S.,Hatanaka
H., Yokoyama S., Tanaka H., Yasuda H., and Shindo H. 2004. NMR
structure of the N-terminal domain of SUMO ligase PIAS1 and its
interaction with tumor suppressor p53 and A/T-rich DNA oligomers. J. B
+iol.Chem. 279: 31455�31461. (18) Leng R.P., Lin Y., Ma W., Wu 
+H., Lemmers B., Chung S., Parant J.M.,
Lozano G., Hakem R., and Benchimol S. 2003. Pirh2, a p53-induced
ubiquitin-protein ligase,promotes p53 degradation. Cell 112: 779&#6553
+3;791. (19) Zhong S.,Salomoni P.,and Pandolfi P.P.2000.The transcript
+ional role
of PML and the nuclear body. Nature Cell Biol. 2: E85�E90. (20)
+ Ito A.,Lai C.H.,Zhao X.,Saito S.,Hamilton M.H.,Appella E.,and Yao
T.P.2001.p300/CBP-mediated p53 acetylation is commonly induced by
p53-activating agents and inhibited by MDM2. EMBO J. 20: 1331�1
+340.
(21) Ando K., Ozaki T.,Yamamoto H., Furuya K., Hosoda M., Hayashi S., 
+Fukuzawa M., and Nakagawara A. 2004. Polo-like kinase 1 (Plk1)
inhibits p53 function by physical interaction and phosphorylation. J. 
+Biol.Chem . 279: 25549�25561. (22) Hanson S., Kim E., and Depp
+ert W. 2005. Redox factor 1 (Ref-1)
enhances specific DNA binding of p53 by promoting p53 tetrameriza-
tion. Oncogene 24: 1641�1647. (23) Jalota A., Singh K., Pavithr
+a L., Kaul-Ghanekar R., Jameel S., and
Chattopadhyay S.2005.Tumor suppressor SMAR1 activates and stabi-
lizes p53 through its arginine-serine-rich motif. J. Biol. Chem. 280: 
+16019�16029.
(24) Markowitz J.,Chen I.,Gitti R.,Baldisseri D.M.,Pan Y.,Udan R.,Carr
+ier
F.,MacKerell A.D.,Jr.,and Weber D.J.2004.Identification and charac-
terization of small molecule inhibitors of the calcium-dependent
S100B-p53 tumor suppressor interaction. J.Med.Chem. 47: 5085�50
+93. (25) Li H.H.,Li A.G.,Sheppard H.M.,and Liu X.2004.Phosphorylation
+ on
Thr-55 by TAF1 mediates degradation ofp53:A role for TAF1 in cell G1
progression. Mol.Cell 13: 867�878. (26) Barlev N.A., Liu L., Ch
+ehab N.H., Mansfield K., Harris K.G., Halazonetis T.D., and Berger S.
+L. 2001. Acetylation of p53 activates
transcription through recruitment of coactivators/histone acetyltrans-
ferases. Mol.Cell 8: 1243�1254. (27) Sommers J.A.,Sharma S.,Doh
+erty K.M.,Karmakar P.,Yang Q.,Kenny
M.K., Harris C.C., and Brosh R.M., Jr. 2005. p53 modulates RPA-
dependent and RPA-independent WRN helicase activity. Cancer Res. 65: 1
+223�1233. (28) Waterman M.J., Stavridi E.S., Waterman J.L., an
+d Halazonetis T.D.
1998. ATM-dependent activation of p53 involves dephosphorylation
and association with 14-3-3 proteins. Nat.Genet. 19: 175�178.
�2005 by Cold Spring Harbor Laboratory Press.All rights reserve
+d.
The reproduction,modification,storage in a retrieval system ,or retran
+smission,in any form or by any means,electronic,mechanical ,or otherw
+ise, for reasons other than personal,noncommercial use is strictly pr
+ohib ited without prior written permission.You are authorized to  dow
+nload one copy of the material on this Web site for personal,noncomme
+rcial use only.The material made available on this Web site is protec
+ted by United States copyright laws and is provided solely for the us
+e of instructors in teaching their courses and assessing student lear
+ning.Dissemination or  sale of any of this materi- al,as a whole or i
+n parts (including the World Wide Web),is not permitted.All users of 
+these materials and visitors to this Web site are expected to abide b
+y these restrictions.Requests for permission for other uses ofthis ma
+t erial should be directed to Cold Spring Harbor Laborator y Press,1 
+Bungtown Road, Cold Spring Harbor,NY11724 or submitted via our World 
+Wide Web Site at http://www.cshlpress.com/.
[download]