GGN3与GGNBP2的相互作用及作用区域确定  

Identification of a Protein Interacting with GGN3 from Mouse Testis cDNA Library by Yeast Two-Hybrid Screening

作　　者：曹志国[1] 张进[1] 周煜[1] 王艳[1] 赵庆国[1] 卢柏松[1] 黄培堂[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2005年第1期5-8,共4页Letters in Biotechnology

摘　　要：生殖细胞缺陷症(gcd)小鼠突变体是上世纪90年代初发现的一种不育突变小鼠,FancL(也叫Pog)的缺失是产生gcd突变小鼠的原因。FANCL是一种含有PHD结构域的泛素E3连接酶,是Fanconi贫血复合物中的组分之一。在生殖细胞中,FANCL与GGN1和GGN3相互作用,而GGN1和GGN3蛋白的功能还不清楚。为了研究GGN3的功能,揭示更多的参与该过程的蛋白质,运用Clontech公司新开发的第三套酵母双杂交系统以GGN3为诱饵从成年小鼠睾丸cDNA文库中筛选与其相互作用的蛋白分子。发现了一个精子生成期间在睾丸中特异性高表达的基因Ggnbp2,免疫共沉淀分析表明,Ggnbp2编码的蛋白质产物GGNBP2在哺乳动物细胞中与GGN3特异相互作用。通过构建突变体,确定了GGNBP2蛋白与GGN3相互作用的区域。以上结果为揭示GGN3和GGNBP2在生殖发育中的功能、丰富生殖细胞发育的蛋白调控网络及其调控规律奠定了一定的基础。The germ cell-deficient (gcd) mutation was discovered in the early of 1990's. The gene underlying this mutation is FancL (also named Pog, proliferation of germ cells). Human FANCL was recently found to be an ubiquitin E3 ligase and a component of the nuclear Fanconi anemia complex. In germ cells, FANCL interacts with GGN1 and GGN3, two proteins from alternative splicing of the same gene Ggn. The function of GGN3 and GGN1 still remains unknown. To screen the protein interacting with GGN3 from mouse testis cDNA library by yeast two-hybrid system, several clones interacting with GGN3 were identified. One of them was homologous with Ggnbp2 (gametogenetin binding protein 2). Co-immunoprecipitation showed that GGNBP2 bound to GGN3 in mammalian cells. Deletion mutation determined the region in GGNBP2 mediating the interaction.

关 键 词：GGN3 GGNBP2 酵母双杂交 免疫共沉淀 相互作用 作用区域 小鼠 生殖细胞缺陷症 FancL 生殖发育 

分 类 号：Q132[生物学—普通生物学] Q78