Efficient 5'-3' DNA end resection by HerA and NurA is essential for cell viability in the crenarchaeon Sulfolobus islandicus.
Efficient 5'-3' DNA end resection by HerA and NurA is essential for cell viability in the crenarchaeon Sulfolobus islandicus.
复制标题
HerA 和 NurA 高效切除 5-3-DNA 末端对于岛状硫化叶菌中的细胞活力至关重要
DOI：
10.1186/s12867-015-0030-z
复制
发表时间：
2015-02-14
影响因子：
--
通讯作者：
Shen Y
中科院分区：
文献类型：
--
作者：
Huang Q;Liu L;Liu J;Ni J;She Q;Shen Y
dna酶/解旋酶和核酸酶在同源重组修复（homologous recombination repair， HRR）中起重要作用。在古细菌中，有关这些酶及其在这一基本而复杂的DNA修复过程中的功能的许多机制细节仍然知之甚少。本研究以嗜热古菌Sulfolobus islandicus为模型，研究了atp酶/解旋酶HerA、核酸酶NurA及其相关蛋白Mre11和Rad50的体内功能。结果通过突变体繁殖实验，我们发现同一个操纵子中的四个基因mre11、rad50、herA和nurA对细胞活力都是必需的。突变蛋白的遗传互补试验与生化表征相结合，表明HerA的atp酶活性、HerA和NurA之间的相互作用以及HerA-NurA复合物的5 ' -3 ' DNA末端切除活性对细胞活力至关重要。在体内用染色体编码的N-His-HerA共纯化了NurA和另外两个可能的HRR蛋白：含有atp酶的PIN （PilT n端）结构域和Holliday连接分解蛋白Hjc。体外拉下进一步证实了HerA与atp酶和Hjc的相互作用。结论HerA-NurA复合物具有高效的5 ' -3 ' DNA末端切除活性，是黄芪中HerA和NurA存在的必要条件，是产生HRR中3 ' -悬垂的关键。除了NurA外，HerA在细胞中可能还有其他结合伙伴。
BackgroundATPase/Helicases and nucleases play important roles in homologous recombination repair (HRR). Many of the mechanistic details relating to these enzymes and their function in this fundamental and complicated DNA repair process remain poorly understood in archaea. Here we employed Sulfolobus islandicus, a hyperthermophilic archaeon, as a model to investigate the in vivo functions of the ATPase/helicase HerA, the nuclease NurA, and their associated proteins Mre11 and Rad50.ResultsWe revealed that each of the four genes in the same operon, mre11, rad50, herA, and nurA, are essential for cell viability by a mutant propagation assay. A genetic complementation assay with mutant proteins was combined with biochemical characterization demonstrating that the ATPase activity of HerA, the interaction between HerA and NurA, and the efficient 5′-3′ DNA end resection activity of the HerA-NurA complex are essential for cell viability. NurA and two other putative HRR proteins: a PIN (PilT N-terminal)-domain containing ATPase and the Holliday junction resolvase Hjc, were co-purified with a chromosomally encoded N-His-HerA in vivo. The interactions of HerA with the ATPase and Hjc were further confirmed by in vitro pull down.ConclusionEfficient 5′-3′ DNA end resection activity of the HerA-NurA complex contributes to necessity of HerA and NurA in Sulfolobus, which is crucial to yield a 3′-overhang in HRR. HerA may have additional binding partners in cells besides NurA.
登录
查看更多内容
影响因子：
3.8
作者：
Hong, Ye;Chu, Mingzhu;Shen, Yulong
通讯作者：
Shen, Yulong
影响因子：
64.5
作者：
Hopkins BB;Paull TT
通讯作者：
Paull TT
影响因子：
4.8
作者：
Fujikane, R;Komori, K;Ishino, Y
通讯作者：
Ishino, Y
影响因子：
2.9
作者：
Contursi, Patrizia;Jensen, Susanne;She, Qunxin
通讯作者：
She, Qunxin
影响因子：
7.7
作者：
Manzan, A;Pfeiffer, G;Hopfner, KP
通讯作者：
Hopfner, KP