Rfc5 in cooperation with Rad24, controls DNA damage checkpoints throughout the cell cycle in Saccharomyces cerevisiae
Rfc5 in cooperation with Rad24, controls DNA damage checkpoints throughout the cell cycle in Saccharomyces cerevisiae
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DOI：
10.1128/mcb.20.16.5888-5896.2000
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发表时间：
2000-08-01
影响因子：
5.3
通讯作者：
Sugimoto, K
中科院分区：
文献类型：
--
作者：
Naiki, T;Shimomura, T;Sugimoto, K
Rad 24和RFC 5是芽殖酵母酿酒酵母中DNA损伤检查点控制所必需的，Rad 24在结构上与复制因子C（RFC）亚基相关，并与RFC亚基Rfc 2、Rfc 3、Rfc 4和Rfc 5缔合。rad24 Δ突变体在所有的G（1）-、S-和G（2）/M-期DNA损伤检查点都有缺陷，而rfc 5 - 1突变体仅在S-期DNA中受损。损坏检查点。RFC亚基和Rad 24都含有三磷酸核苷（NTP）结合的共有序列。为了确定NTP结合基序是否对Rad 24功能重要，我们突变了该基序中保守的赖氨酸（115）残基。rad 24-K115 E突变（将赖氨酸变为谷氨酸）导致完全功能丧失表型，而rad 24-K115 R突变（将赖氨酸变为精氨酸）则没有表现出明显的表型。尽管rfc 5 -1和rad 24-K115 R单突变体在G（1）和G（2）/M期DNA损伤检查点上都没有缺陷，但rfc 5 -1 rad 24-K115 R双突变体在这些检查点上变得有缺陷。免疫共沉淀实验表明Rad 24（K115 R）不能与rfc 5-l突变体中的RFC蛋白相互作用。总之，这些结果表明，RFC 5，像RAD 24一样，在所有的G（1）-，S-和G（2）/M-期DNA损伤检查点中发挥作用，并表明Rad 24与RFC蛋白的相互作用对于DNA损伤检查点控制是必不可少的。
RAD24 and RFC5 are required for DNA damage checkpoint control in the budding yeast Saccharomyces cerevisiae, Rad24 is structurally related to replication factor C (RFC) subunits and associates with RFC subunits Rfc2, Rfc3, Rfc4, and Rfc5. rad24 Delta. mutants are defective in all the G(1)-, S-, and G(2)/M-phase DNA damage checkpoints, whereas the rfc5-l mutant is impaired only in the S-phase DNA. damage checkpoint. Both the RFC subunits and Rad24 contain a consensus sequence for nucleoside triphosphate (NTP) binding. To determine whether the NTP-binding motif is important for Rad24 function, we mutated the conserved lysine(115) residue in this motif. The rad24-K115E mutation, which changes lysine to glutamate, confers a complete loss-of-function phenotype, while the rad24-K115R mutation, which changes lysine to arginine, shows no apparent phenotype. Although neither rfc5-1 nor rad24-K115R single mutants are defective in the G(1)- and G(2)/M-phase DNA damage checkpoints, rfc5-1 rad24-K115R double mutants become defective in these checkpoints. Coimmunoprecipitation experiments revealed that Rad24(K115R) fails to interact with the RFC proteins in rfc5-l mutants. Together, these results indicate that RFC5, like RAD24, functions in all the G(1)-, S- and G(2)/M-phase DNA damage checkpoints and suggest that the interaction of Rad24 with the RFC proteins is essential for DNA damage checkpoint control.