Wen Y D, Perissi V, Staszewski L M, Yang W M, Krones A, Glass C K, Rosenfeld M G, Seto E
Department of Chemistry, H. Lee Moffitt Cancer Center and Research Institute, University of South Florida, Tampa, FL 33612, USA.
Proc Natl Acad Sci U S A. 2000 Jun 20;97(13):7202-7. doi: 10.1073/pnas.97.13.7202.
Acetylation and deacetylation of nucleosomal histones have profound effects on gene transcription in all eukaryotes. In humans, three highly homologous class I and four class II histone deacetylase (HDAC) enzymes have been identified to date. The class I deacetylases HDAC1 and HDAC2 are components of multisubunit complexes, one of which could associate with the nuclear hormone receptor corepressor, N-CoR. N-CoR also interacts with class II deacetylases HDAC4, HDAC5, and HDAC7. In comparison with HDAC1 and HDAC2, HDAC3 remains relatively uncharacterized, and very few proteins have been shown to interact with HDAC3. Using an affinity purification approach, we isolated an enzymatically active HDAC3 complex that contained members of the nuclear receptor corepressor family. Deletion analysis of N-CoR revealed that HDAC3 binds multiple N-CoR regions in vitro and that all of these regions are required for maximal binding in vivo. The N-CoR domains that interact with HDAC3 are distinct from those that bind other HDACs. Transient overexpression of HDAC3 and microinjection of Abs against HDAC3 showed that a component of transcriptional repression mediated by N-CoR depends on HDAC3. Interestingly, data suggest that interaction with a region of N-CoR augments the deacetylase activity of HDAC3. These results provide a possible molecular mechanism for HDAC3 regulation and argue that N-CoR is a platform in which distinct domains can interact with most of the known HDACs.
核小体组蛋白的乙酰化和去乙酰化对所有真核生物的基因转录都有深远影响。在人类中,迄今已鉴定出三种高度同源的I类和四种II类组蛋白去乙酰化酶(HDAC)。I类去乙酰化酶HDAC1和HDAC2是多亚基复合物的组成部分,其中之一可与核激素受体共抑制因子N-CoR结合。N-CoR还与II类去乙酰化酶HDAC4、HDAC5和HDAC7相互作用。与HDAC1和HDAC2相比,HDAC3的特征相对较少,并且很少有蛋白质被证明与HDAC3相互作用。我们采用亲和纯化方法,分离出一种含有核受体共抑制因子家族成员的具有酶活性的HDAC3复合物。对N-CoR的缺失分析表明,HDAC3在体外结合多个N-CoR区域,且所有这些区域在体内实现最大结合都是必需的。与HDAC3相互作用的N-CoR结构域与那些结合其他HDAC的结构域不同。HDAC3的瞬时过表达以及针对HDAC3的抗体的显微注射表明,由N-CoR介导的转录抑制的一个组成部分依赖于HDAC3。有趣的是,数据表明与N-CoR的一个区域相互作用可增强HDAC3的去乙酰化酶活性。这些结果为HDAC3的调控提供了一种可能的分子机制,并表明N-CoR是一个平台,在这个平台中不同的结构域可与大多数已知的HDAC相互作用。