A role for CENP-A/Cse4 phosphorylation on serine 33 in deposition at the centromere
A role for CENP-A/Cse4 phosphorylation on serine 33 in deposition at the centromere
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DOI：
10.1093/femsyr/fox094
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发表时间：
2018-02-01
影响因子：
3.2
通讯作者：
Ehrenhofer-Murray, Ann E.
中科院分区：
文献类型：
--
作者：
Hoffmann, Gesine;Samel-Pommerencke, Anke;Ehrenhofer-Murray, Ann E.
着丝粒是着丝粒的组装位点，在细胞分裂过程中，着丝粒将染色单体连接到微管以分离姐妹染色单体。着丝粒的特征在于存在组蛋白H3变体CENP-A（在酿酒酵母中称为Cse 4）。本文研究了Cse 4的丝氨酸33磷酸化（Cse 4-S33 ph）在S.酿酒酵母，其位于延伸的Cse 4 N-末端的必需N-末端结构域（END）内。值得注意的是，我们鉴定了组蛋白H4-K5，8，12 R导致温度敏感性生长缺陷，Cse 4-S33突变以及对诺考达唑和羟基脲的敏感性。此外，Cse 4-S33 ph的缺乏降低了Cse 4在着丝粒序列的水平，表明Cse 4沉积在S33磷酸化的缺乏下是有缺陷的。我们还鉴定了与组蛋白H2 A-E57 A和H2 A-L 66 A的合成遗传相互作用，这两者都导致与组蛋白伴侣FACT的相互作用减少和染色质中H2 A/H2 B水平降低，再次支持H2 A/H2 B和Cse 4沉积的组合缺陷导致着丝粒缺陷的观点。总之，我们的数据突出了正确的组蛋白沉积在构建功能性着丝粒核小体中的重要性，并表明Cse 4-S33 ph在此过程中的作用。
Centromeres are the sites of assembly of the kinetochore, which connect the chromatids to the microtubules for sister chromatid segregation during cell division. Centromeres are characterized by the presence of the histone H3 variant CENP-A (termed Cse4 in Saccharomyces cerevisiae). Here, we investigated the function of serine 33 phosphorylation of Cse4 (Cse4-S33ph) in S. cerevisiae, which lies within the essential N-terminal domain (END) of the extended Cse4 N-terminus. Significantly, we identified histone H4-K5, 8, 12R to cause a temperature-sensitive growth defect with mutations in Cse4-S33 and sensitivity to nocodazole and hydroxyurea. Furthermore, the absence of Cse4-S33ph reduced the levels of Cse4 at centromeric sequences, suggesting that Cse4 deposition is defective in the absence of S33 phosphorylation. We furthermore identified synthetic genetic interactions with histone H2A-E57A and H2A-L66A, which both cause a reduced interaction with the histone chaperone FACT and reduced H2A/H2B levels in chromatin, again supporting the notion that a combined defect of H2A/H2B and Cse4 deposition causes centromeric defects. Altogether, our data highlight the importance of correct histone deposition in building a functional centromeric nucleosome and suggests a role for Cse4-S33ph in this process.