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首页> 外文期刊>Molecular and Cellular Biology >A Novel Role for the Mitotic Spindle during DNA Segregation in Yeast: Promoting 2μm Plasmid-Cohesin Association
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A Novel Role for the Mitotic Spindle during DNA Segregation in Yeast: Promoting 2μm Plasmid-Cohesin Association

机译：酵母中DNA分离过程中有丝分裂纺锤体的新型作用：促进2μm质粒-Cohesin协会。

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The 2μm circle plasmid in Saccharomyces cerevisiae is a model for a stable, high-copy-number, extrachromosomal “selfish” DNA element. By combining a partitioning system and an amplification system, the plasmid ensures its stable propagation and copy number maintenance, even though it does not provide any selective advantage to its host. Recent evidence suggests that the partitioning system couples plasmid segregation to chromosome segregation. We now demonstrate an unexpected and unconventional role for the mitotic spindle in the plasmid-partitioning pathway. The spindle specifies the nuclear address of the 2μm circle and promotes recruitment of the cohesin complex to the plasmid-partitioning locus STB. Only the nuclear microtubules, and not the cytoplasmic ones, are required for loading cohesin at STB. In cells recovering from nocodazole-induced spindle depolymerization and G2/M arrest, cohesin-STB association can be established coincident with spindle restoration. This postreplication recruitment of cohesin is not functional in equipartitioning. However, normally acquired cohesin can be inactivated after replication without causing plasmid missegregation. In the mtw1-1 mutant yeast strain, the plasmid cosegregates with the spindle and the spindle-associated chromosomes; by contrast, a substantial number of the chromosomes are not associated with the spindle. These results are consistent with a model in which the spindle promotes plasmid segregation in a chromosome-linked fashion.
机译：酿酒酵母（Saccharomyces cerevisiae）中的2μm环状质粒是稳定，高拷贝数，染色体外“自私” DNA元件的模型。通过结合分配系统和扩增系统，质粒可确保其稳定的繁殖和拷贝数维持，即使它没有为其宿主提供任何选择优势。最近的证据表明该分配系统将质粒分离与染色体分离耦合在一起。我们现在证明有丝分裂纺锤体在质粒分配途径中的意外和非常规作用。纺锤体指定2μm圈的核地址，并促进粘着蛋白复合物募集到质粒分配位点 STB 。在 STB 上加载粘着蛋白仅需要核微管，而不需要细胞质。在从诺考达唑诱导的纺锤体解聚和G 2 / M阻滞中恢复的细胞中，可与纺锤体恢复同时建立粘着蛋白- STB 缔合。复制后的粘着蛋白募集在均分中不起作用。但是，正常情况下获得的粘着蛋白可以在复制后被灭活，而不会引起质粒错聚。在 mtw1 - 1 突变酵母菌株中，质粒与纺锤体和纺锤体相关染色体共分离。相反，大量的染色体与纺锤不相关。这些结果与其中纺锤体以染色体连锁的方式促进质粒分离的模型一致。

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