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Cohesin ATPase activities regulate DNA binding and coiled-coil configuration
Proceedings of the National Academy of Sciences of the United States of America ( IF 11.1 ) Pub Date : 2022-08-08 , DOI: 10.1073/pnas.2208004119
Xingya Xu 1 , Ryuta Kanai 2 , Li Wang 1 , Mitsuhiro Yanagida 1
Affiliation  

The cohesin complex is required for sister chromatid cohesion and genome compaction. Cohesin coiled coils (CCs) can fold at break sites near midpoints to bring head and hinge domains, located at opposite ends of coiled coils, into proximity. Whether ATPase activities in the head play a role in this conformational change is yet to be known. Here, we dissected functions of cohesin ATPase activities in cohesin dynamics in Schizosaccharomyces pombe . Isolation and characterization of cohesin ATPase temperature-sensitive (ts) mutants indicate that both ATPase domains are required for proper chromosome segregation. Unbiased screening of spontaneous suppressor mutations rescuing the temperature lethality of cohesin ATPase mutants identified several suppressor hotspots in cohesin that located outside of ATPase domains. Then, we performed comprehensive saturation mutagenesis targeted to these suppressor hotspots. Large numbers of the identified suppressor mutations indicated several different ways to compensate for the ATPase mutants: 1) Substitutions to amino acids with smaller side chains in coiled coils at break sites around midpoints may enable folding and extension of coiled coils more easily; 2) substitutions to arginine in the DNA binding region of the head may enhance DNA binding; or 3) substitutions to hydrophobic amino acids in coiled coils, connecting the head and interacting with other subunits, may alter conformation of coiled coils close to the head. These results reflect serial structural changes in cohesin driven by its ATPase activities potentially for packaging DNAs.

中文翻译:

Cohesin ATPase 活性调节 DNA 结合和卷曲螺旋结构

cohesin 复合物是姐妹染色单体凝聚和基因组压缩所必需的。Cohesin 螺旋线圈 (CC) 可以在中点附近的断裂点折叠,使位于螺旋线圈相对两端的头部和铰链域靠近。头部的 ATPase 活动是否在这种构象变化中起作用尚不清楚。在这里,我们剖析了 cohesin ATPase 活动在 cohesin 动力学中的功能粟酒裂殖酵母. cohesin ATPase 温度敏感 (ts) 突变体的分离和表征表明,正确的染色体分离需要两个 ATPase 结构域。对自发抑制突变的无偏筛选挽救了黏连蛋白 ATPase 突变体的温度致死性,确定了位于 ATPase 结构域外的黏连蛋白中的几个抑制热点。然后,我们针对这些抑制热点进行了全面的饱和诱变。大量已识别的抑制突变表明有几种不同的方式来补偿 ATPase 突变体:1)在中点周围的断裂位点处用较小侧链的氨基酸替代卷曲螺旋可能使卷曲螺旋更容易折叠和延伸;2) 在头部的 DNA 结合区域替换为精氨酸可能会增强 DNA 结合;或 3) 取代卷曲螺旋中的疏水性氨基酸,连接头部并与其他亚基相互作用,可能会改变靠近头部的卷曲螺旋的构象。这些结果反映了 cohesin 的一系列结构变化,这些变化是由其可能用于包装 DNA 的 ATPase 活性驱动的。
更新日期:2022-08-08
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