摘要
PCAF(P300/CBP associated factor,P300/CBP相关因子)是一种重要的组蛋白乙酰转移酶,主要通过使核小体组蛋白发生乙酰化而参与基因转录的调控。P300/CBP是一种能够与PCAF相互作用的宿主细胞蛋白,定位于细胞核ND10结构(nuclear dot10)。ND10是由70余种蛋白质分子组成的、以分散的斑点或团块样方式存在于细胞核内的大分子复合物,与基因转录调控过程有着密切关系。早幼粒细胞白血病蛋白(promyelocytic leukemia protein,PML)是ND10的主要组成成分,对于维持ND10的正常结构与功能至关重要。在成功构建pGBK-PML3、pGAD-PCAF、pGEX-PCAF、pFlag-PCAF和pcDNA-PML3重组质粒的基础上,用酵母双杂交证明PCAF与PML3之间存在着相互作用;用GST-pull down技术证明GST-PCAF能够将PML3蛋白捕获并沉降下来,表明在体外条件下两者之间存在直接的相互作用;用免疫共沉淀和荧光共定位证明在体内PCAF能够与PML发生直接的相互作用而定位于ND10。这些结果为进一步深入研究PCAF的生物学功能和作用模式奠定了基础。
As one of the important histone acetyltransferases,PCAF（P300/CBP associated factor）mainly mediates the acetylation of core histones and hence plays a vital role in the transcriptional regulation.Several cellular proteins that interact with PCAF directly,such as P300/CBP,p53 and MDM2,have been reported localized in nuclear dot 10（ND10）,an intranuclear structure with a typical discrete punctuate distribution.ND10 contains more than 70 kinds of proteins and has been implicated in the regulation of gene transcription.The most key component of ND10 is PML（promyelocytic leukemia protein）.PML is well known to be essential for the maintenance of the normal structure and function of ND10.By the use of yeast two-hybridization,GST pull-down,co-immunoprecipitation and fluorescence colocalization assays,the evidence for the direct interaction of PCAF with PML and the localization of PCAF in ND10 was provided,which will be beneficial for the further research on the biological function of PCAF.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2010年第6期21-27,共7页
China Biotechnology
基金
河南省自然科学基金(623031300)
河南省教育厅自然科学研究(2009A310009)资助项目
