应用酵母双杂交技术寻找与GGNBP相互作用的蛋白质

Searching for GGNBP interacting proteins using yeast two-hybrid system

生殖细胞缺陷症(gcd)小鼠突变体是20个世纪90年代初发现的一种不育突变小鼠,其不育原因是由于其胚胎期原始生殖细胞的数目低于正常。FancL(也叫Pog)的缺失是引起gcd突变小鼠的原因,FancL基因缺失后可能影响了小鼠胚胎期原始生殖细胞的增殖/存活和成年期小鼠精母细胞的减数分裂。FANCL是一种含有PHD结构域的泛素E3连接酶,是Fanconi贫血复合物的组分之一。在生殖细胞中,FANCL与GGN1和GGN3相互作用,GGN1和GGN2又与一种新的蛋白质GGNBP特异作用。但GGNBP蛋白的功能还不清楚。为了研究GGNBP的功能以及揭示更多的参与该过程的蛋白质,运用Clontech公司新开发的第3套酵母双杂交系统,以GGNBP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质基因,发现了一个主要在睾丸中表达的新的基因,其编码的蛋白质产物在酵母系统中与GGNBP特异作用。

The germ cell-deficient(gcd)mutation was discovered in the early of1990's.The phenotype of gcd mutant,characterized by adult sterility,is due to a deficiency of primordial germ cells(PGC)during the embryonic stage.The gene underlying this mutation is FancL(also named Pog),which seemed to affect the proliferation/survival of PGC during the embryonic stage and the meiosis of spermatocytes in the adulthood.Human FANCL was recently found to be an ubiquitin E3ligase and a component of the nuclear Fanconi anemia complex.In germ cells,FANCL interacts with GGN1and GGN3,two proteins from alternative splicing of the same gene Ggn.GGN1and GGN2interact with another novel protein GGNBP,the function of which still remains unknown.In this study,the yeast two-hybrid system is used to search for pro-teins interacting with GGNBP.In the yeast system,GGNBP is founed to interact with a novel unnamed protein whose function remains to be clarified.