G0 Cell Unit, Okinawa Institute of Science and Technology Graduate University.
Institute of Future Agriculture, Northwest A&F University, Yangling, Shaanxi, People's Republic of China.
Proc Jpn Acad Ser B Phys Biol Sci. 2023;99(3):61-74. doi: 10.2183/pjab.99.005.
Cohesin is a heteropentameric protein complex that contributes to various aspects of chromosome structure and function, such as sister chromatid cohesion, genome compaction, and DNA damage response. Previous studies have provided abundant information on architecture and regional structures of the cohesin complex, but the configuration and structural dynamics of the whole cohesin complex are still largely unknown, partly due to flexibility of its coiled coils. We studied cohesin organization and dynamics using in vivo functional mutation compensation. Specifically, we developed and applied genetic suppressor screening methods to identify second mutations in cohesin complex genes that rescue lethality caused by various site-specific abnormalities in the cohesin complex. Functional analysis of these missense suppressor mutations revealed novel features of cohesin. Here, we summarize recent genetic suppressor screening results and insights into: 1) cohesin's structural organization when holding chromosomal DNAs; 2) interaction between cohesin head-kleisin and hinge; 3) ATP-driven cohesin conformational changes for genome packaging.
着丝粒蛋白是一种异五聚体蛋白复合物,有助于染色体结构和功能的各个方面,如姐妹染色单体黏合、基因组紧缩和 DNA 损伤反应。先前的研究提供了大量关于着丝粒复合物结构和区域结构的信息,但着丝粒复合物的整体构象和结构动力学在很大程度上仍然未知,部分原因是其螺旋线圈的灵活性。我们使用体内功能突变补偿研究了着丝粒的组织和动态。具体来说,我们开发并应用了遗传抑制筛选方法来鉴定着丝粒复合物基因中的第二突变,这些突变可以挽救着丝粒复合物中各种特定位置异常引起的致死性。对这些错义抑制突变的功能分析揭示了着丝粒的新特征。在这里,我们总结了最近的遗传抑制筛选结果和对以下方面的见解:1)当结合染色体 DNA 时着丝粒的结构组织;2)着丝粒头部-黏连蛋白和铰链之间的相互作用;3)用于基因组包装的 ATP 驱动的着丝粒构象变化。