当前位置: X-MOL 学术PLOS Genet. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Skp, Cullin, F-box (SCF)-Met30 and SCF-Cdc4-Mediated Proteolysis of CENP-A Prevents Mislocalization of CENP-A for Chromosomal Stability in Budding Yeast.
PLOS Genetics ( IF 4.0 ) Pub Date : 2020-02-07 , DOI: 10.1371/journal.pgen.1008597
Wei-Chun Au 1 , Tianyi Zhang 1 , Prashant K Mishra 1 , Jessica R Eisenstatt 1 , Robert L Walker 1 , Josefina Ocampo 2 , Anthony Dawson 1 , Jack Warren 1 , Michael Costanzo 3 , Anastasia Baryshnikova 4 , Karin Flick 5 , David J Clark 2 , Paul S Meltzer 1 , Richard E Baker 6 , Chad Myers 7 , Charles Boone 3 , Peter Kaiser 5 , Munira A Basrai 1
Affiliation  

Restricting the localization of the histone H3 variant CENP-A (Cse4 in yeast, CID in flies) to centromeres is essential for faithful chromosome segregation. Mislocalization of CENP-A leads to chromosomal instability (CIN) in yeast, fly and human cells. Overexpression and mislocalization of CENP-A has been observed in many cancers and this correlates with increased invasiveness and poor prognosis. Yet genes that regulate CENP-A levels and localization under physiological conditions have not been defined. In this study we used a genome-wide genetic screen to identify essential genes required for Cse4 homeostasis to prevent its mislocalization for chromosomal stability. We show that two Skp, Cullin, F-box (SCF) ubiquitin ligases with the evolutionarily conserved F-box proteins Met30 and Cdc4 interact and cooperatively regulate proteolysis of endogenous Cse4 and prevent its mislocalization for faithful chromosome segregation under physiological conditions. The interaction of Met30 with Cdc4 is independent of the D domain, which is essential for their homodimerization and ubiquitination of other substrates. The requirement for both Cdc4 and Met30 for ubiquitination is specifc for Cse4; and a common substrate for Cdc4 and Met30 has not previously been described. Met30 is necessary for the interaction between Cdc4 and Cse4, and defects in this interaction lead to stabilization and mislocalization of Cse4, which in turn contributes to CIN. We provide the first direct link between Cse4 mislocalization to defects in kinetochore structure and show that SCF-mediated proteolysis of Cse4 is a major mechanism that prevents stable maintenance of Cse4 at non-centromeric regions, thus ensuring faithful chromosome segregation. In summary, we have identified essential pathways that regulate cellular levels of endogenous Cse4 and shown that proteolysis of Cse4 by SCF-Met30/Cdc4 prevents mislocalization and CIN in unperturbed cells.

中文翻译:

Skp,Cullin,F-box(SCF)-Met30和SCF-Cdc4介导的CENP-A的蛋白水解可防止CENP-A错位定位以提高酵母芽中的染色体稳定性。

将组蛋白H3变体CENP-A(酵母中的Cse4,果蝇中的CID)的定位限制到着丝粒对于忠实的染色体分离至关重要。CENP-A的错误定位会导致酵母,果蝇和人类细胞中的染色体不稳定(CIN)。在许多癌症中都观察到CENP-A的过度表达和定位错误,这与浸润性增加和预后不良有关。尚未定义在生理条件下调节CENP-A水平和定位的基因。在这项研究中,我们使用了全基因组的遗传筛选来鉴定Cse4稳态所需的必要基因,以防止其对染色体稳定性的错误定位。我们展示了两个Skp,Cullin,F-box(SCF)泛素连接酶与进化上保守的F-box蛋白Met30和Cdc4相互作用并共同调节内源性Cse4的蛋白水解,并防止其在生理条件下忠实染色体分离的错误定位。Met30与Cdc4的相互作用不依赖于D结构域,D结构域对于它们的同质二聚化和其他底物的泛素化至关重要。泛素化对Cdc4和Met30的要求都针对Cse4。并且以前没有描述用于Cdc4和Met30的通用底物。Met30对于Cdc4和Cse4之间的相互作用是必需的,并且此相互作用中的缺陷导致Cse4的稳定化和错位,进而导致CIN。我们提供了Cse4错定位与动线粒结构缺陷之间的第一个直接联系,并表明SCF介导的Cse4蛋白水解是防止Cse4在非着丝粒区域稳定维持的主要机制,从而确保了忠实的染色体分离。总而言之,我们已经确定了调节内源性Cse4细胞水平的基本途径,并表明SCF-Met30 / Cdc4对Cse4的蛋白水解可防止未受干扰的细胞发生错位和CIN。
更新日期:2020-03-05
down
wechat
bug