Cheng Hwei-Ling, Mostoslavsky Raul, Saito Shin'ichi, Manis John P, Gu Yansong, Patel Parin, Bronson Roderick, Appella Ettore, Alt Frederick W, Chua Katrin F
Howard Hughes Medical Institute, Children's Hospital, Center for Blood Research, and Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA.
Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):10794-9. doi: 10.1073/pnas.1934713100. Epub 2003 Sep 5.
SIRT1 is a mammalian homolog of the Saccharomyces cerevisiae chromatin silencing factor Sir2. Dominant-negative and overexpression studies have implicated a role for SIRT1 in deacetylating the p53 tumor suppressor protein to dampen apoptotic and cellular senescence pathways. To elucidate SIRT1 function in normal cells, we used gene-targeted mutation to generate mice that express either a mutant SIRT1 protein that lacks part of the catalytic domain or has no detectable SIRT1 protein at all. Both types of SIRT1 mutant mice and cells had essentially the same phenotypes. SIRT1 mutant mice were small, and exhibited notable developmental defects of the retina and heart, and only infrequently survived postnatally. Moreover, SIRT1-deficient cells exhibited p53 hyperacetylation after DNA damage and increased ionizing radiation-induced thymocyte apoptosis. In SIRT1-deficient embryonic fibroblasts, however, p53 hyperacetylation after DNA damage was not accompanied by increased p21 protein induction or DNA damage sensitivity. Together, our observations provide direct evidence that endogenous SIRT1 protein regulates p53 acetylation and p53-dependent apoptosis, and show that the function of this enzyme is required for specific developmental processes.
SIRT1是酿酒酵母染色质沉默因子Sir2在哺乳动物中的同源物。显性负性和过表达研究表明,SIRT1在使p53肿瘤抑制蛋白去乙酰化以抑制凋亡和细胞衰老途径中发挥作用。为了阐明SIRT1在正常细胞中的功能,我们利用基因靶向突变生成了两种小鼠,一种表达缺乏部分催化结构域的突变型SIRT1蛋白,另一种完全没有可检测到的SIRT1蛋白。这两种类型的SIRT1突变小鼠和细胞具有基本相同的表型。SIRT1突变小鼠体型较小,表现出明显的视网膜和心脏发育缺陷,出生后很少存活。此外,SIRT1缺陷细胞在DNA损伤后表现出p53高度乙酰化,并且电离辐射诱导的胸腺细胞凋亡增加。然而,在SIRT1缺陷的胚胎成纤维细胞中,DNA损伤后p53高度乙酰化并未伴随着p21蛋白诱导增加或DNA损伤敏感性增加。总之,我们的观察结果提供了直接证据,证明内源性SIRT1蛋白调节p53乙酰化和p53依赖性凋亡,并表明该酶的功能是特定发育过程所必需的。