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首页> 外文期刊>Molecular and Cellular Biology >Rfc4 Interacts with Rpa1 and Is Required for Both DNA Replication and DNA Damage Checkpoints in Saccharomyces cerevisiae
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Rfc4 Interacts with Rpa1 and Is Required for Both DNA Replication and DNA Damage Checkpoints in Saccharomyces cerevisiae

机译：Rfc4与Rpa1相互作用，是酿酒酵母中DNA复制和DNA损伤检查点所必需的

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The large subunit of replication protein A (Rpa1) consists of three single-stranded DNA binding domains and an N-terminal domain (Rpa1N) of unknown function. To determine the essential role of this domain we searched for mutations that require wild-type Rpa1N for viability in yeast. A mutation in RFC4, encoding a small subunit of replication factor C (RFC), was found to display allele-specific interactions with mutations in the gene encoding Rpa1 (RFA1). Mutations that map to Rpa1N and confer sensitivity to the DNA synthesis inhibitor hydroxyurea, such asrfa1-t11, are lethal in combination withrfc4-2. The rfc4-2 mutant itself is sensitive to hydroxyurea, and like rfc2 and rfc5 strains, it exhibits defects in the DNA replication block and intra-S checkpoints. RFC4 and the DNA damage checkpoint geneRAD24 were found to be epistatic with respect to DNA damage sensitivity. We show that the rfc4-2 mutant is defective in the G1/S DNA damage checkpoint response and that both therfc4-2 and rfa1-t11 strains are defective in the G2/M DNA damage checkpoint. Thus, in addition to its essential role as part of the clamp loader in DNA replication, Rfc4 plays a role as a sensor in multiple DNA checkpoint pathways. Our results suggest that a physical interaction between Rfc4 and Rpa1N is required for both roles.
机译：复制蛋白A的大亚基（Rpa1）由三个单链DNA结合结构域和一个功能未知的N末端结构域（Rpa1N）组成。为了确定该结构域的重要作用，我们搜索了需要野生型Rpa1N才能在酵母中存活的突变。发现 RFC4 中的一个编码复制因子C（RFC）小亚基的突变与编码Rpa1（ RFA1 ）的基因中的突变显示等位基因特异性相互作用。映射到Rpa1N并赋予DNA合成抑制剂羟基脲敏感性的突变，例如 rfa1-t11 ，与 rfc4-2 结合使用具有致命性。 rfc4-2 突变体本身对羟基脲敏感，并且与 rfc2 和 rfc5 菌株一样，它在DNA复制区和内部片段中均存在缺陷。 S个检查站。发现 RFC4 和DNA损伤检查点基因 RAD24 在DNA损伤敏感性方面具有上位性。我们显示 rfc4-2 突变体在G 1 / S DNA损伤检查点响应中有缺陷，并且 rfc4-2 和< em> rfa1-t11 菌株在G 2 / M DNA损伤检查点中存在缺陷。因此，Rfc4除了在DNA复制中作为钳位装载器的一部分起着至关重要的作用外，还在多个DNA检查点途径中作为传感器发挥作用。我们的结果表明，Rfc4和Rpa1N之间的物理相互作用对于这两个角色都是必需的。

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