The ctf13-30/CTF13 genomic haploinsufficiency modifier screen identifies the yeast chromatin remodeling complex RSC, which is required for the establishment of sister chromatid cohesion
The ctf13-30/CTF13 genomic haploinsufficiency modifier screen identifies the yeast chromatin remodeling complex RSC, which is required for the establishment of sister chromatid cohesion
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DOI：
10.1128/mcb.24.3.1232-1244.2003
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发表时间：
2004-02-01
影响因子：
5.3
通讯作者：
Hieter, P
中科院分区：
文献类型：
--
作者：
Baetz, KK;Krogan, NJ;Hieter, P
芽殖酵母着丝粒-动粒复合体通过介导染色体沿着纺锤体微管的附着和运动，确保有丝分裂和减数分裂中染色体的高保真分离。为了识别新的基因和途径，其功能影响染色体传递，我们开发了一个基因组单倍不足修饰筛选，并使用ctf 13 -30，编码突变的核心动粒蛋白，作为参考点。我们证明，通过一系列的二次筛选，基因组修饰剂筛选是一种成功的方法，用于识别编码非必需蛋白质的基因，所需的染色体分离的保真度。在我们的筛选中分离的一个基因是RSC 2，RSC染色质重塑复合物的非必需亚基。RSC 2突变体在染色体分离和内聚方面都有缺陷，但动粒蛋白在着丝粒上的定位不受影响。我们确定，在没有RSC 2的情况下，粘着蛋白仍然可以与染色体结合，但不能在姐妹染色单体之间实现适当的凝聚力，这表明RSC在凝聚力的建立中起作用。此外，许多RSC的亚基进行了亲和纯化，并确定了RSC的一个新的组成部分，Rtt 102。我们的工作表明，只有一个子集的非必需的RSC亚基的功能，在维持染色体的传输保真度。
The budding yeast centromere-kinetochore complex ensures high-fidelity chromosome segregation in mitosis and meiosis by mediating the attachment and movement of chromosomes along spindle microtubulles. To identify new genes and pathways whose function impinges on chromosome transmission, we developed a genomic haploinsufficiency modifier screen and used ctf13-30, encoding a mutant core kinetochore protein, as the reference point. We demonstrate through a series of secondary screens that the genomic modifier screen is a successful method for identifying genes that encode nonessential proteins required for the fidelity of chromosome segregation. One gene isolated in our screen was RSC2, a nonessential subunit of the RSC chromatin remodeling complex. rsc2 mutants have defects in both chromosome segregation and cohesion, but the localization of kinetochore proteins to centromeres is not affected. We determined that, in the absence of RSC2, cohesin could still associate with chromosomes but fails to achieve proper cohesion between sister chromatids, indicating that RSC has a role in the establishment of cohesion. In addition, numerous subunits of RSC were affinity purified and a new component of RSC, Rtt102, was identified. Our work indicates that only a subset of the nonessential RSC subunits function in maintaining chromosome transmission fidelity.