SOLO在果蝇减数分裂黏连中的作用机理

黏连（cohesion）由黏连体（cohesin）介导，是减数分裂中染色体正确分离必不可少的。减数分裂黏连体及其亚基REC8在其它一些生物中均有深入研究，但黑腹果蝇减数分裂黏连体及REC8至今没有得到阐明。我们前期鉴定的减数分裂特异蛋白SOLO可能是果蝇的黏连体亚基，可能替代REC8的功能。本项目拟定点突变SOLO潜在Separase切割位点，运用遗传分析，免疫荧光，免疫电泳等方法研究Separase潜在位点突变对减数分裂黏连和染色体分离以及对Separase切割的影响；拟定点突变SOLO的C-末端保守氨基酸，分析其突变对SOLO与黏连体亚基SMC1的相互作用，以及对SMC1在染色体上定位及减数分裂黏连的影响；结合免疫共沉淀和质谱等方法确定SOLO所在黏连体的亚基组成。通过研究，将揭示SOLO在果蝇黏连中的作用机理，丰富对减数分裂黏连体的认识，从而对减数分裂研究领域产生重大和深远的影响。

Meiotic cohesion is mediated by cohesin and is essential for proper chromosome segregation in meiosis. Meiotic cohesin and its subunit REC8 have been studied heavily in other organisms, however, the meiotic cohesin and REC8 in Drosophila have not been clarified yet. We previously identified a meiosis-specific protein SOLO that may be a subunit of the Drosophila meiotic cohesin, replacing the functions of REC8. In the project the potential Separase cleavage sites of SOLO will be mutated by site-directed mutagenesis and the effects of the mutations on meiotic cohesion, chromosome segregation and Separase cleavage will be studied. The C-terminal conserved residues of SOLO will be mutated by site-directed mutagenesis to explore the effects of the mutations on the interactions between SOLO and SMC1, and on SMC1’s chromosomal location and meiotic cohesion. The composition of meiotic cohesin (s) containing SOLO in Drosophila will be determined by combining co-immunoprecipitation and mass spectrometry. Our study will reveal the working mechanism of SOLO in Drosophila meiotic cohesion, enrich the knowledge of meiotic cohesin, therefore, making significant and profound impacts in the field of meiotic research.