Ctf19p: A novel kinetochore protein in Saccharomyces cerevisiae and a potential link between the kinetochore and mitotic spindle.
Ctf19p: A novel kinetochore protein in Saccharomyces cerevisiae and a potential link between the kinetochore and mitotic spindle.
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DOI：
10.1083/jcb.145.1.15
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发表时间：
1999-04-05
期刊：
影响因子：
--
通讯作者：
Hieter P
中科院分区：
文献类型：
--
作者：
Hyland KM;Kingsbury J;Koshland D;Hieter P
利用编码已知着丝点蛋白的CTF13作为过表达内参基因进行遗传合成剂量致死（SDL）筛选，鉴定出两个染色体传递保真度（ctf）突变体YCTF58和YCTF26。这些突变株携带一种新基因的独立等位基因，我们将其命名为CTF19。鉴于其在着丝点功能中的潜在作用，我们对CTF19基因进行了详细的克隆和表征。CTF19编码一种非必需的369个氨基酸的蛋白质。ctf19突变株表现出严重的染色体错分离表型，对苯甲酰过敏，并在循环细胞中以G2/M积累。CTF19基因上与着丝粒结构突变体和有丝分裂检查点突变体相互作用。此外，在体外实验中，ctf19突变体在小染色体上的着丝粒结合微管的能力上表现出缺陷。体内交联和染色质免疫沉淀表明Ctf19p与CEN DNA特异性相互作用。此外，Ctf19-HAp定位于纺锤体极体的核面，并与纺锤体相关蛋白发生遗传相互作用。我们认为Ctf19p是一个大分子着丝点复合体的一部分，它可能是着丝点和有丝分裂纺锤体之间的联系。
A genetic synthetic dosage lethality (SDL) screen using CTF13 encoding a known kinetochore protein as the overexpressed reference gene identified two chromosome transmission fidelity (ctf) mutants, YCTF58 and YCTF26. These mutant strains carry independent alleles of a novel gene, which we have designated CTF19. In light of its potential role in kinetochore function, we have cloned and characterized the CTF19 gene in detail. CTF19 encodes a nonessential 369–amino acid protein. ctf19 mutant strains display a severe chromosome missegregation phenotype, are hypersensitive to benomyl, and accumulate at G2/M in cycling cells. CTF19 genetically interacts with kinetochore structural mutants and mitotic checkpoint mutants. In addition, ctf19 mutants show a defect in the ability of centromeres on minichromosomes to bind microtubules in an in vitro assay. In vivo cross-linking and chromatin immunoprecipitation demonstrates that Ctf19p specifically interacts with CEN DNA. Furthermore, Ctf19-HAp localizes to the nuclear face of the spindle pole body and genetically interacts with a spindle-associated protein. We propose that Ctf19p is part of a macromolecular kinetochore complex, which may func- tion as a link between the kinetochore and the mitotic spindle.
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影响因子：
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