Inhibition of SIRT1 catalytic activity increases p53 acetylation but does not alter cell survival following DNA damage

被引:395
作者
Solomon, JM [1 ]
Pasupuleti, R [1 ]
Xu, L [1 ]
McDonagh, T [1 ]
Curtis, R [1 ]
DiStefano, PS [1 ]
Huber, LJ [1 ]
机构
[1] Elixir Pharmaceut Inc, Cambridge, MA 02139 USA
关键词
D O I
10.1128/MCB.26.1.28-38.2006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Human SIRT1 is an enzyme that deacetyllates the p53 tumor suppressor protein and has been suggested to modulate p53-dependent functions including DNA damage-induced cell death. In this report, we used EX-527, a novel, potent, and specific small-molecule inhibitor of SIRT1 catalytic activity to examine the role of SIRT1 in p53 acetylation and cell survival after DNA damage. Treatment with EX-527 dramatically increased acetylation at lysine 382 of p53 after different types of DNA damage in primary human mammary epithelial cells and several cell lines. Significantly, inhibition of SIRT1 catalytic activity by EX-527 had no effect on cell growth, viability, or p53-controlled gene expression in cells treated with etoposide. Acetyl-p53 was also increased by the histone deacetylase (HDAC) class I/II inhibitor trichostatin A (TSA). EX-527 and TSA acted synergistically to increase acetyl-p53 levels, confirming that p53 acetyllation is regulated by both SIRT1 and HDACs. While TSA alone reduced cell survival after DNA damage, the combination of EX-527 and TSA had no further effect on cell viability and growth. These results show that, although SIRT1 deacetylates p53, this does not play a role in cell survival following DNA damage in certain cell lines and primary human mammary epithelial cells.
引用
收藏
页码:28 / 38
页数:11
相关论文
共 56 条
[1]   Post-translational modifications and activation of p53 by genotoxic stresses [J].
Appella, E ;
Anderson, CW .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 2001, 268 (10) :2764-2772
[2]   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
[3]   Inhibition of silencing and accelerated aging by nicotinamide, a putative negative regulator of yeast Sir2 and human SIRT1 [J].
Bitterman, KJ ;
Anderson, RM ;
Cohen, HY ;
Latorre-Esteves, M ;
Sinclair, DA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (47) :45099-45107
[4]   The Sir2 family of protein deacetylases [J].
Blander, G ;
Guarente, L .
ANNUAL REVIEW OF BIOCHEMISTRY, 2004, 73 :417-435
[5]   Calorie restriction, SIRT1 and metabolism: Understanding longevity [J].
Bordone, L ;
Guarente, L .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2005, 6 (04) :298-305
[6]   SIRT1 deacetylation and repression of p300 involves lysine residues 1020/1024 within the cell cycle regulatory domain 1 [J].
Bouras, T ;
Fu, MF ;
Sauve, AA ;
Wang, F ;
Quong, AA ;
Perkins, ND ;
Hay, RT ;
Gu, W ;
Pestell, RG .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (11) :10264-10276
[7]   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
[8]   Stress-dependent regulation of FOXO transcription factors by the SIRT1 deacetylase [J].
Brunet, A ;
Sweeney, LB ;
Sturgill, JF ;
Chua, KF ;
Greer, PL ;
Lin, YX ;
Tran, H ;
Ross, SE ;
Mostoslavsky, R ;
Cohen, HY ;
Hu, LS ;
Cheng, HL ;
Jedrychowski, MP ;
Gygi, SP ;
Sinclair, DA ;
Alt, FW ;
Greenberg, ME .
SCIENCE, 2004, 303 (5666) :2011-2015
[9]   Restoration of the tumor suppressor function to mutant p53 by a low-molecular-weight compound [J].
Bykov, VJN ;
Issaeva, N ;
Shilov, A ;
Hultcrantz, M ;
Pugacheva, E ;
Chumakov, P ;
Bergman, J ;
Wiman, KG ;
Selivanova, G .
NATURE MEDICINE, 2002, 8 (03) :282-288
[10]   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