Mck1-mediated proteolysis of CENP-A prevents mislocalization of CENP-A for chromosomal stability in Saccharomyces cerevisiae.

IF 3.3 3区生物学 Q2 GENETICS & HEREDITY

Genetics Pub Date : 2024-09-04 DOI:10.1093/genetics/iyae108

Tianyi Zhang, Wei-Chun Au, Kentaro Ohkuni, Roshan L Shrestha, Peter Kaiser, Munira A Basrai

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引用次数: 0

Abstract

Centromeric localization of evolutionarily conserved CENP-A (Cse4 in Saccharomyces cerevisiae) is essential for chromosomal stability. Mislocalization of overexpressed CENP-A to noncentromeric regions contributes to chromosomal instability in yeasts, flies, and humans. Overexpression and mislocalization of CENP-A observed in many cancers are associated with poor prognosis. Previous studies have shown that F-box proteins, Cdc4 and Met30 of the Skp, Cullin, F-box ubiquitin ligase cooperatively regulate proteolysis of Cse4 to prevent Cse4 mislocalization and chromosomal instability under normal physiological conditions. Mck1-mediated phosphorylation of Skp, Cullin, F-box-Cdc4 substrates such as Cdc6 and Rcn1 enhances the interaction of the substrates with Cdc4. Here, we report that Mck1 interacts with Cse4, and Mck1-mediated proteolysis of Cse4 prevents Cse4 mislocalization for chromosomal stability. Our results showed that mck1Δ strain overexpressing CSE4 (GAL-CSE4) exhibits lethality, defects in ubiquitin-mediated proteolysis of Cse4, mislocalization of Cse4, and reduced Cse4-Cdc4 interaction. Strain expressing GAL-cse4-3A with mutations in three potential Mck1 phosphorylation consensus sites (S10, S16, and T166) also exhibits growth defects, increased stability with mislocalization of Cse4-3A, chromosomal instability, and reduced interaction with Cdc4. Constitutive expression of histone H3 (Δ16H3) suppresses the chromosomal instability phenotype of GAL-cse4-3A strain, suggesting that the chromosomal instability phenotype is linked to Cse4-3A mislocalization. We conclude that Mck1 and its three potential phosphorylation sites on Cse4 promote Cse4-Cdc4 interaction and this contributes to ubiquitin-mediated proteolysis of Cse4 preventing its mislocalization and chromosomal instability. These studies advance our understanding of pathways that regulate cellular levels of CENP-A to prevent mislocalization of CENP-A in human cancers.

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在酿酒酵母中，Mck1 介导的 CENP-A 蛋白水解可防止 CENP-A 误定位，从而提高染色体稳定性。

进化保守的 CENP-A（酿酒酵母中的 Cse4）的中心粒定位对染色体稳定性至关重要。在酵母、苍蝇和人类中，过表达的 CENP-A 在非中心粒区域的错定位导致染色体不稳定（CIN）。在许多癌症中观察到的 CENP-A 过表达和错定位与预后不良有关。先前的研究表明，在正常生理条件下，F-盒蛋白、Cdc4 和 Skp、Cullin、F-盒（SCF）泛素连接酶的 Met30 可协同调节 Cse4 的蛋白水解，防止 Cse4 错定位和 CIN。Mck1 介导的 SCF-Cdc4 底物（如 Cdc6 和 Rcn1）磷酸化可增强底物与 Cdc4 的相互作用。在这里，我们报告了Mck1与Cse4的相互作用，以及Mck1介导的Cse4蛋白水解阻止了Cse4为染色体稳定而发生的错定位。我们的研究结果表明，过表达CSE4（GAL-CSE4）的mck1Δ菌株表现出致死性、泛素介导的Cse4蛋白水解缺陷、Cse4错定位和Cse4-Cdc4相互作用减少。表达 GAL-cse4-3A 并在三个潜在的 Mck1 磷酸化共识位点（S10、S16 和 T166）发生突变的菌株也表现出生长缺陷、稳定性增加、Cse4-3A 和 CIN 定位错误以及与 Cdc4 的相互作用减少。组蛋白 H3（Δ16H3）的连续表达抑制了 GAL-cse4-3A 株系的 CIN 表型，表明 CIN 表型与 Cse4-3A 的错定位有关。我们的结论是，Mck1及其在Cse4上的三个潜在磷酸化位点促进了Cse4-Cdc4的相互作用，这有助于泛素介导的Cse4蛋白水解，防止其误定位和CIN。这些研究加深了我们对调节细胞中 CENP-A 水平的途径的了解，从而防止 CENP-A 在人类癌症中的误定位。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Genetics GENETICS & HEREDITY-

CiteScore

6.90

自引率

6.10%

发文量

177

审稿时长

1.5 months

期刊介绍： GENETICS is published by the Genetics Society of America, a scholarly society that seeks to deepen our understanding of the living world by advancing our understanding of genetics. Since 1916, GENETICS has published high-quality, original research presenting novel findings bearing on genetics and genomics. The journal publishes empirical studies of organisms ranging from microbes to humans, as well as theoretical work. While it has an illustrious history, GENETICS has changed along with the communities it serves: it is not your mentor''s journal. The editors make decisions quickly – in around 30 days – without sacrificing the excellence and scholarship for which the journal has long been known. GENETICS is a peer reviewed, peer-edited journal, with an international reach and increasing visibility and impact. All editorial decisions are made through collaboration of at least two editors who are practicing scientists. GENETICS is constantly innovating: expanded types of content include Reviews, Commentary (current issues of interest to geneticists), Perspectives (historical), Primers (to introduce primary literature into the classroom), Toolbox Reviews, plus YeastBook, FlyBook, and WormBook (coming spring 2016). For particularly time-sensitive results, we publish Communications. As part of our mission to serve our communities, we''ve published thematic collections, including Genomic Selection, Multiparental Populations, Mouse Collaborative Cross, and the Genetics of Sex.