Ohkuni Kentaro, Abdulle Rashid, Tong Amy Hin Yan, Boone Charles, Kitagawa Katsumi
Department of Molecular Pharmacology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
PLoS One. 2008 Feb 20;3(2):e1617. doi: 10.1371/journal.pone.0001617.
The spindle checkpoint ensures the accurate segregation of chromosomes by monitoring the status of kinetochore attachment to microtubules. Simultaneous mutations in one of several kinetochore and cohesion genes and a spindle checkpoint gene cause a synthetic-lethal or synthetic-sick phenotype. A synthetic genetic array (SGA) analysis using a mad2Delta query mutant strain of yeast identified YBP2, a gene whose product shares sequence similarity with the product of YBP1, which is required for H(2)O(2)-induced oxidation of the transcription factor Yap1. ybp2Delta was sensitive to benomyl and accumulated at the mitotic stage of the cell cycle. Ybp2 physically associates with proteins of the COMA complex (Ctf19, Okp1, Mcm21, and Ame1) and 3 components of the Ndc80 complex (Ndc80, Nuf2, and Spc25 but not Spc24) in the central kinetochore and with Cse4 (the centromeric histone and CENP-A homolog). Chromatin-immunoprecipitation analyses revealed that Ybp2 associates specifically with CEN DNA. Furthermore, ybp2Delta showed synthetic-sick interactions with mutants of the genes that encode the COMA complex components. Ybp2 seems to be part of a macromolecular kinetochore complex and appears to contribute to the proper associations among the central kinetochore subcomplexes and the kinetochore-specific nucleosome.
纺锤体检查点通过监测动粒与微管的附着状态来确保染色体的准确分离。几个动粒和黏连蛋白基因之一与一个纺锤体检查点基因同时发生突变会导致合成致死或合成病态表型。使用酵母的mad2Δ查询突变菌株进行的合成遗传阵列(SGA)分析鉴定出YBP2,该基因的产物与YBP1的产物具有序列相似性,YBP1是H₂O₂诱导转录因子Yap1氧化所必需的。ybp2Δ对苯菌灵敏感,并在细胞周期的有丝分裂阶段积累。Ybp2在中央动粒中与COMA复合体(Ctf19、Okp1、Mcm21和Ame1)的蛋白以及Ndc80复合体的3个组分(Ndc80、Nuf2和Spc25,但不包括Spc24)发生物理关联,并与Cse4(着丝粒组蛋白和CENP - A同源物)发生物理关联。染色质免疫沉淀分析表明,Ybp2特异性地与CEN DNA结合。此外,ybp2Δ与编码COMA复合体组分的基因突变体表现出合成病态相互作用。Ybp2似乎是一个大分子动粒复合体的一部分,并且似乎有助于中央动粒亚复合体与动粒特异性核小体之间的正确关联。