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首页> 外文期刊>Nucleic Acids Research >Genetic and functional interactions between Mus81-Mms4 and Rad27
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Genetic and functional interactions between Mus81-Mms4 and Rad27

机译：Mus81-Mms4和Rad27之间的遗传和功能相互作用

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摘要

The two endonucleases, Rad27 (yeast Fen1) and Dna2, jointly participate in the processing of Okazaki fragments in yeasts. Mus81-Mms4 is a structure-specific endonuclease that can resolve stalled replication forks as well as toxic recombination intermediates. In this study, we show that Mus81-Mms4 can suppress dna2 mutational defects by virtue of its functional and physical interaction with Rad27. Mus81-Mms4 stimulated Rad27 activity significantly, accounting for its ability to restore the growth defects caused by the dna2 mutation. Interestingly, Rad27 stimulated the rate of Mus81-Mms4 catalyzed cleavage of various substrates, including regressed replication fork substrates. The ability of Rad27 to stimulate Mus81-Mms4 did not depend on the catalytic activity of Rad27, but required the C-terminal 64 amino acid fragment of Rad27. This indicates that the stimulation was mediated by a specific protein-protein interaction between the two proteins. Our in vitro data indicate that Mus81-Mms4 and Rad27 act together during DNA replication and resolve various structures that can impede normal DNA replication. This conclusion was further strengthened by the fact that rad27 mus81 or rad27 mms4 double mutants were synergistically lethal. We discuss the significance of the interactions between Rad27, Dna2 and Mus81-Mms4 in context of DNA replication.
机译：Rad27（酵母Fen1）和Dna2这两个核酸内切酶共同参与酵母中冈崎片段的加工。 Mus81-Mms4是一种结构特异性核酸内切酶，可以解决停滞的复制叉以及有毒重组中间体。在这项研究中，我们表明Mus81-Mms4可以通过与Rad27的功能和物理相互作用来抑制dna2突变缺陷。 Mus81-Mms4显着刺激了Rad27的活性，这说明其能够恢复由dna2突变引起的生长缺陷的能力。有趣的是，Rad27刺激了Mus81-Mms4催化的各种底物（包括回归复制叉底物）裂解的速率。 Rad27刺激Mus81-Mms4的能力不取决于Rad27的催化活性，但需要Rad27的C端64个氨基酸片段。这表明刺激是由两种蛋白质之间特定的蛋白质-蛋白质相互作用介导的。我们的体外数据表明，Mus81-Mms4和Rad27在DNA复制过程中共同起作用，并解析出各种可能阻碍正常DNA复制的结构。 rad27 mus81或rad27 mms4双重突变具有协同致命性，这一事实进一步加强了这一结论。我们讨论了在DNA复制的背景下Rad27，Dna2和Mus81-Mms4之间相互作用的重要性。

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