基础医学与临床 ›› 2008, Vol. 28 ›› Issue (6): 529-534.

• 研究论文 •    下一篇

NSPc1相互作用蛋白的筛查及其与组蛋白乙酰化酶HBO1相互作用的验证

龚燕华 石磊 吴旭东 强伯勤 彭小忠 袁建刚   

  1. 基础所医学分子生物学国家重点实验室 北京协和医学院医学分子生物学国家重点实验室 北京协和医学院医学分子生物学国家重点实验室 北京协和医学院医学分子生物学国家重点实验室 基础所分子生物学系 基础所分子生物学系
  • 收稿日期:2007-12-28 修回日期:2008-02-28 出版日期:2008-06-25 发布日期:2008-06-25
  • 通讯作者: 龚燕华

Screening and verification of the interactive proteins for nervous system polycomb 1

Yan-hua GONG, Lei SHI, Xu-dong WU, Bo-qin QIANG, Xiao-zhong PENG, Jian-gang YUAN   

  1. National Laboratory of Medical Molecular Biology,Institute of Basic Medical Sciences,CAMS & PUMC
  • Received:2007-12-28 Revised:2008-02-28 Online:2008-06-25 Published:2008-06-25
  • Contact: Yan-hua GONG,

摘要: 目的 寻找多梳家族(Polycomb group)蛋白NSPc1的体内可能的相互作用蛋白。 方法 通过酵母双杂交实验筛选与NSPc1相互作用的蛋白。使用Pull-down实验,Co-IP实验以及细胞内荧光共定位实验进一步证实双杂交中发现的相互作用。结果 分别以NSPc1蛋白的全长、N端和C端作为诱饵筛查人3月胎脑cDNA文库,发现N端诱饵可以与组蛋白乙酰化转移酶HBO1相互作用,该相互作用在体内外实验中都得到了验证。结论 组蛋白乙酰化酶HBO1是NSPc1的相互作用蛋白之一,该相互作用可能参与NSPc1对靶基因转录的表观抑制活性的发挥。

Abstract: Objective To find out the protein complex members associated with the nervous system polycomb 1 (NSPc1) in vivo. Methods Use the yeast two hybrid system to screen the potential interactive proteins of NSPc1, and verify the interaction by GST pull-down, co-immunoprecipitation and subcellular co-localization assays. Results The full length, N-terminus and C-terminus of NSPc1 were used as the "baits" to screen the 3-month fetal brain cDNA library in yeast. Some proteins, including HBO1 (Histone Acetyltransferase Binding to ORC-1), were found to be interactive proteins of NSPc1, then the interaction between HBO1 and NSPc1 was verified both in vitro and in vivo. Conclusion The histone acetyltransferase HBO1 is one of the interactive proteins of NSPc1, and the interaction may be involved in the epigenetic repression activity of polycomb proteins.