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Swi1 and Swi3 Are Components of a Replication Fork Protection Complex in Fission Yeast

机译：Swi1和Swi3是裂变酵母中复制叉保护复合体的组件

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Swi1 is required for programmed pausing of replication forks near the mat1 locus in the fission yeast Schizosaccharomyces pombe. This fork pausing is required to initiate a recombination event that switches mating type. Swi1 is also needed for the replication checkpoint that arrests division in response to fork arrest. How Swi1 accomplishes these tasks is unknown. Here we report that Swi1 copurifies with a 181-amino-acid protein encoded by swi3+. The Swi1-Swi3 complex is required for survival of fork arrest and for activation of the replication checkpoint kinase Cds1. Association of Swi1 and Swi3 with chromatin during DNA replication correlated with movement of the replication fork. swi1Δ and swi3Δ mutants accumulated Rad22 (Rad52 homolog) DNA repair foci during replication. These foci correlated with the Rad22-dependent appearance of Holliday junction (HJ)-like structures in cells lacking Mus81-Eme1 HJ resolvase. Rhp51 and Rhp54 homologous recombination proteins were not required for viability in swi1Δ or swi3Δ cells, indicating that the HJ-like structures arise from single-strand DNA gaps or rearranged forks instead of broken forks. We propose that Swi1 and Swi3 define a fork protection complex that coordinates leading- and lagging-strand synthesis and stabilizes stalled replication forks.
机译：在分裂酵母 Schizosaccharomyces pombe 的 mat1 位点附近，有计划地暂停复制叉的程序需要Swi1。要启动切换配对类型的重组事件，需要此分叉暂停。复制检查点也需要Swi1，该检查点可响应分叉逮捕而逮捕部门。 Swi1如何完成这些任务是未知的。在这里，我们报道Swi1与 swi3 + 编码的181个氨基酸共纯化。 Swi1-Swi3复合物对于叉式逮捕的存活和复制检查点激酶Cds1的激活是必需的。 DNA复制过程中Swi1和Swi3与染色质的关联与复制叉的移动相关。 s wi1Δ和swi3Δ突变体在复制过程中积累了Rad22（Rad52同源物）DNA修复灶。这些病灶与缺少Mus81-Eme1 HJ分解酶的细胞中霍利迪结（HJ）样结构的Rad22依赖的外观相关。 Rhm51和Rhp54同源重组蛋白对于swi1Δ或swi3Δ细胞的生存力不是必需的，这表明HJ样结构是由单链DNA缺口或重新排列的叉子而非断的叉子。我们建议Swi1和Swi3定义一个分叉保护复合体，该复合体可协调前导链和滞后链合成并稳定停滞的复制分叉。

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