Genome instability due to ribonucleotide incorporation into DNA.
Genome instability due to ribonucleotide incorporation into DNA.
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DOI：
10.1038/nchembio.424
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发表时间：
2010-10
影响因子：
14.8
通讯作者：
Kunkel, Thomas A.
中科院分区：
文献类型：
--
作者：
McElhinny, Stephanie A. Nick;Kumar, Dinesh;Clark, Alan B.;Watt, Danielle L.;Watts, Brian E.;Lundstrom, Else-Britt;Johansson, Erik;Chabes, Andrei;Kunkel, Thomas A.
维持DNA的化学特性依赖于DNA聚合酶对核糖核苷酸的排除。然而，体外DNA合成过程中核糖核苷酸的排除尚不完善。为了确定核糖核苷酸是否在体内DNA复制过程中被纳入，我们用亮氨酸或甘氨酸取代了酵母DNA聚合酶ε (Pol ε)的活性位点蛋氨酸。与野生型Pol ε相比，M644L Pol ε的核糖核苷酸结合率低3倍，而M644G Pol ε的核糖核苷酸结合率高11倍。这种层次结构在缺乏RNase H2的酵母菌株中被重新屈服。此外，pol2-M644G rnh201Δ菌株在s期进展较慢，dNTP池升高，在重复序列中以高速率和基因取向依赖的方式产生2-5个碱基对缺失。这些数据表明，核糖核苷酸在体内复制过程中被纳入，它们被依赖于RNase h2的修复去除，修复缺陷通过DNA链错位导致复制应激和基因组不稳定。
Maintaining the chemical identity of DNA depends on ribonucleotide exclusion by DNA polymerases. However, ribonucleotide exclusion during DNA synthesis in vitro is imperfect. To determine if ribonucleotides are incorporated during DNA replication in vivo, we substituted leucine or glycine for an active site methionine in yeast DNA polymerase ε (Pol ε). Compared to wild type Pol ε, ribonucleotide incorporation in vitro was 3-fold lower for M644L and 11-fold higher for M644G Pol ε. This hierarchy was re-capitulated in vivo in yeast strains lacking RNase H2. Moreover, the pol2-M644G rnh201Δ strain progressed more slowly through S-phase, had elevated dNTP pools and generated 2–5 base pair deletions in repetitive sequences at a high rate and gene orientation-dependent manner. The data indicate that ribonucleotides are incorporated during replication in vivo, that they are removed by RNase H2-dependent repair, and that defective repair results in replicative stress and genome instability via DNA strand misalignment.
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影响因子：
5.3
作者：
KUNKEL, TA;HAMATAKE, RK;SUGINO, A
通讯作者：
SUGINO, A
影响因子：
5.6
作者：
Bonnin, A;Lázaro, JM;Salas, M
通讯作者：
Salas, M
DOI：
10.1073/pnas.94.5.1619
发表时间：
1997-03-04
影响因子：
11.1
作者：
Joyce, CM
通讯作者：
Joyce, CM
影响因子：
64.5
作者：
Chabes, A;Georgieva, B;Thelander, L
通讯作者：
Thelander, L
影响因子：
30.8
作者：
Crow, Yanick J.;Leitch, Andrea;Jackson, Andrew P.
通讯作者：
Jackson, Andrew P.