Unligated Okazaki Fragments Induce PCNA Ubiquitination and a Requirement for Rad59-Dependent Replication Fork Progression.
Unligated Okazaki Fragments Induce PCNA Ubiquitination and a Requirement for Rad59-Dependent Replication Fork Progression.
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DOI：
10.1371/journal.pone.0066379
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Bielinsky AK
中科院分区：
文献类型：
--
作者：
Nguyen HD;Becker J;Thu YM;Costanzo M;Koch EN;Smith S;Myung K;Myers CL;Boone C;Bielinsky AK
在芽殖酵母中由CDC 9编码的DNA连接酶I的缺乏导致未连接的冈崎片段的积累，并在非典型赖氨酸残基处触发PCNA泛素化。该信号对于激活S期检查点至关重要，S期检查点促进细胞周期延迟。我们在这里报告说，pol 30-K107突变减轻cdc 9突变体细胞周期延迟，与K107的PCNA修饰影响复制叉处DNA合成速率的想法一致。为了确定PCNA泛素化是否发生在响应缺口或由PCNA-DNA连接酶相互作用的缺乏引发，我们补充cdc 9细胞与野生型DNA连接酶I或突变形式，这不能与PCNA相互作用。这两种酶逆转了PCNA泛素化，认为这种修饰可能是一种新的切口感觉机制的组成部分，而不是由于在cdc 9突变体中自发发生的非特异性继发突变。为了进一步了解细胞如何科普DNA复制过程中切口的积累，我们在全基因组合成致死性筛选中利用cdc 9 -1，该筛选将RAD 59鉴定为强负相互作用物。与cdc 9单突变体相比，cdc 9 rad 59 Δ双突变体没有改变PCNA泛素化，但增强了复制检查点Mrc 1介导物的磷酸化。由于Mrc 1驻留在复制叉上，并响应于叉停滞而被磷酸化，这些结果表明Rad 59抑制了切口诱导的复制叉减慢。因此，我们建议Rad 59促进叉进展时，冈崎片段加工受到损害，并抵消PCNA-K107介导的细胞周期阻滞。
Deficiency in DNA ligase I, encoded by CDC9 in budding yeast, leads to the accumulation of unligated Okazaki fragments and triggers PCNA ubiquitination at a non-canonical lysine residue. This signal is crucial to activate the S phase checkpoint, which promotes cell cycle delay. We report here that a pol30-K107 mutation alleviated cell cycle delay in cdc9 mutants, consistent with the idea that the modification of PCNA at K107 affects the rate of DNA synthesis at replication forks. To determine whether PCNA ubiquitination occurred in response to nicks or was triggered by the lack of PCNA-DNA ligase interaction, we complemented cdc9 cells with either wild-type DNA ligase I or a mutant form, which fails to interact with PCNA. Both enzymes reversed PCNA ubiquitination, arguing that the modification is likely an integral part of a novel nick-sensory mechanism and not due to non-specific secondary mutations that could have occurred spontaneously in cdc9 mutants. To further understand how cells cope with the accumulation of nicks during DNA replication, we utilized cdc9-1 in a genome-wide synthetic lethality screen, which identified RAD59 as a strong negative interactor. In comparison to cdc9 single mutants, cdc9 rad59Δ double mutants did not alter PCNA ubiquitination but enhanced phosphorylation of the mediator of the replication checkpoint, Mrc1. Since Mrc1 resides at the replication fork and is phosphorylated in response to fork stalling, these results indicate that Rad59 alleviates nick-induced replication fork slowdown. Thus, we propose that Rad59 promotes fork progression when Okazaki fragment processing is compromised and counteracts PCNA-K107 mediated cell cycle arrest.
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在基因组量表上对酵母中适应性和遗传相互作用的定量分析。
影响因子：
48
作者：
Baryshnikova, Anastasia;Costanzo, Michael;Kim, Yungil;Ding, Huiming;Koh, Judice;Toufighi, Kiana;Youn, Ji-Young;Ou, Jiongwen;San Luis, Bryan-Joseph;Bandyopadhyay, Sunayan;Hibbs, Matthew;Hess, David;Gingras, Anne-Claude;Bader, Gary D.;Troyanskaya, Olga G.;Brown, Grant W.;Andrews, Brenda;Boone, Charles;Myers, Chad L.
通讯作者：
Myers, Chad L.
影响因子：
5.3
作者：
Guzder, SN;Sommers, CH;Prakash, S
通讯作者：
Prakash, S
DOI：
10.1126/science.1180823
发表时间：
2010-01-22
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Costanzo M;Baryshnikova A;Bellay J;Kim Y;Spear ED;Sevier CS;Ding H;Koh JL;Toufighi K;Mostafavi S;Prinz J;St Onge RP;VanderSluis B;Makhnevych T;Vizeacoumar FJ;Alizadeh S;Bahr S;Brost RL;Chen Y;Cokol M;Deshpande R;Li Z;Lin ZY;Liang W;Marback M;Paw J;San Luis BJ;Shuteriqi E;Tong AH;van Dyk N;Wallace IM;Whitney JA;Weirauch MT;Zhong G;Zhu H;Houry WA;Brudno M;Ragibizadeh S;Papp B;Pál C;Roth FP;Giaever G;Nislow C;Troyanskaya OG;Bussey H;Bader GD;Gingras AC;Morris QD;Kim PM;Kaiser CA;Myers CL;Andrews BJ;Boone C
通讯作者：
Boone C
影响因子：
56.9
作者：
Groth, Anja;Corpet, Armelle;Almouzni, Genevieve
通讯作者：
Almouzni, Genevieve
影响因子：
21.3
作者：
Das-Bradoo, Sapna;Nguyen, Hai Dang;Bielinsky, Anja-Katrin
通讯作者：
Bielinsky, Anja-Katrin