Acetylation of p53 augments its site-specific DNA binding both in vitro and in vivo

被引:336
作者
Luo, JY
Li, MY
Tang, Y
Laszkowska, M
Roeder, RG
Gu, W
机构
[1] Columbia Univ Coll Phys & Surg, Inst Canc Genet, New York, NY 10032 USA
[2] Columbia Univ Coll Phys & Surg, Dept Pathol, New York, NY 10032 USA
[3] Rockefeller Univ, Biochem & Mol Biol Lab, New York, NY 10021 USA
关键词
ChlP; transcription; CBP/p300;
D O I
10.1073/pnas.0308762101
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
p53 promotes tumor suppression through its ability to function as a transcriptional factor and is activated by posttranslational modifications that include acetylation. Our earlier study demonstrated that p53 acetylation can enhance its sequence-specific DNA binding in vitro, and this notion was later confirmed in several other studies. However, a recent study has reported that in vitro acetylation of p53 fails to stimulate its DNA binding to large DNA fragments, raising an important issue that requires further investigation. Here, we show that unacetylated p53 is able to bind weakly to its consensus site within the context of large DNA fragments, although it completely fails to bind the same site within short oligonucleotide probes. Strikingly, by using highly purified and fully acetylated p53 proteins obtained from cells, we show that acetylation of the C-terminal domain can dramatically enhance site-specific DNA binding on both short oligonucleotide probes and long DNA fragments. Moreover, endogenous p53 apparently can be fully acetylated in response to DNA damage when both histone deacetylase complex 1 (HDAC1)- and Sir2-mediated deacetylation are inhibited, indicating dynamic p53 acetylation and deacetylation events during the DNA damage response. Finally, we also show that acetylation of endogenous p53 indeed significantly augments its ability to bind an endogenous target gene and that p53 acetylation levels correlate well with p53-mediated transcriptional activation in vivo. Thus, our results clarify some of the confusion surrounding acetylation-mediated effects on p53 binding to DNA and suggest that acetylation of p53 in vivo may contribute, at least in part, to its transcriptional activation functions.
引用
收藏
页码:2259 / 2264
页数:6
相关论文
共 60 条
[1]  
Appella E, 2000, PATHOL BIOL, V48, P227
[2]   Recruitment of p300/CBP in p53-dependent signal pathways [J].
Avantaggiati, ML ;
Ogryzko, V ;
Gardner, K ;
Giordano, A ;
Levine, AS ;
Kelly, K .
CELL, 1997, 89 (07) :1175-1184
[3]   MDM2 EXPRESSION IS INDUCED BY WILD TYPE-P53 ACTIVITY [J].
BARAK, Y ;
JUVEN, T ;
HAFFNER, R ;
OREN, M .
EMBO JOURNAL, 1993, 12 (02) :461-468
[4]   Acetylation of p53 activates transcription through recruitment of coactivators/histone acetyltransferases [J].
Barlev, NA ;
Liu, L ;
Chehab, NH ;
Mansfield, K ;
Harris, KG ;
Halazonetis, TD ;
Berger, SL .
MOLECULAR CELL, 2001, 8 (06) :1243-1254
[5]   Ubiquitination, phosphorylation and acetylation: the molecular basis for p53 regulation [J].
Brooks, CL ;
Gu, W .
CURRENT OPINION IN CELL BIOLOGY, 2003, 15 (02) :164-171
[6]   Developmental defects and p53 hyperacetylation in Sir2 homolog (SIRT1)-deficient mice [J].
Cheng, HL ;
Mostoslavsky, R ;
Saito, S ;
Manis, JP ;
Gu, YS ;
Patel, P ;
Bronson, R ;
Appella, E ;
Alt, FW ;
Chua, KF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (19) :10794-10799
[7]   DNA damage-dependent acetylation of p73 dictates the selective activation of apoptotic target genes [J].
Costanzo, A ;
Merlo, P ;
Pediconi, N ;
Fulco, M ;
Sartorelli, V ;
Cole, PA ;
Fontemaggi, G ;
Fanciulli, M ;
Schiltz, L ;
Blandino, G ;
Balsano, C ;
Levrero, M .
MOLECULAR CELL, 2002, 9 (01) :175-186
[8]   TRANSCRIPTIONAL ACTIVATION BY P53 CORRELATES WITH SUPPRESSION OF GROWTH BUT NOT TRANSFORMATION [J].
CROOK, T ;
MARSTON, NJ ;
SARA, EA ;
VOUSDEN, KH .
CELL, 1994, 79 (05) :817-827
[9]   WAF1, A POTENTIAL MEDIATOR OF P53 TUMOR SUPPRESSION [J].
ELDEIRY, WS ;
TOKINO, T ;
VELCULESCU, VE ;
LEVY, DB ;
PARSONS, R ;
TRENT, JM ;
LIN, D ;
MERCER, WE ;
KINZLER, KW ;
VOGELSTEIN, B .
CELL, 1993, 75 (04) :817-825
[10]   Transcriptional regulation by p53 through intrinsic DNA/chromatin binding and site-directed cofactor recruitment [J].
Espinosa, JM ;
Emerson, BM .
MOLECULAR CELL, 2001, 8 (01) :57-69