The signature motif of the Saccharomyces cerevisiae Pif1 DNA helicase is essential in vivo for mitochondrial and nuclear functions and in vitro for ATPase activity.
The signature motif of the Saccharomyces cerevisiae Pif1 DNA helicase is essential in vivo for mitochondrial and nuclear functions and in vitro for ATPase activity.
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DOI：
10.1093/nar/gky655
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发表时间：
2018-09-19
影响因子：
14.9
通讯作者：
Zakian VA
中科院分区：
文献类型：
--
作者：
Geronimo CL;Singh SP;Galletto R;Zakian VA
Pif 1家族DNA解旋酶从细菌到人类都是保守的，并且在体内具有促进基因组完整性的关键和多样的功能。Pif 1家族解旋酶共有一个23个氨基酸的区域，称为Pif 1签名基序（SM），这是该家族所独有的。为了确定SM的重要性，我们对酿酒酵母Pif 1（ScPif 1）的SM进行了突变和功能分析。这些突变删除了SM的部分，使SM中的一个或多个单个氨基酸发生变化，用来自细菌Pif 1家族解旋酶的对应物取代SM，并用来自另一种解旋酶的α-螺旋结构域取代SM中形成α螺旋的部分。对突变体进行了线粒体DNA的维持、端粒和双链断裂处端粒酶的抑制以及冈崎片段成熟的促进测试。虽然SM中某些单一氨基酸的变化是可以耐受的，但ScPif 1 SM的存在和序列对于所有测试的体内功能都是必不可少的。与体内分析一致，体外研究表明，ScPif 1 SM的存在和序列对ATP酶活性至关重要，但对底物结合无关。
Pif1 family DNA helicases are conserved from bacteria to humans and have critical and diverse functions in vivo that promote genome integrity. Pif1 family helicases share a 23 amino acid region, called the Pif1 signature motif (SM) that is unique to this family. To determine the importance of the SM, we did mutational and functional analysis of the SM from the Saccharomyces cerevisiae Pif1 (ScPif1). The mutations deleted portions of the SM, made one or multiple single amino acid changes in the SM, replaced the SM with its counterpart from a bacterial Pif1 family helicase and substituted an α-helical domain from another helicase for the part of the SM that forms an α helix. Mutants were tested for maintenance of mitochondrial DNA, inhibition of telomerase at telomeres and double strand breaks, and promotion of Okazaki fragment maturation. Although certain single amino acid changes in the SM can be tolerated, the presence and sequence of the ScPif1 SM were essential for all tested in vivo functions. Consistent with the in vivo analyses, in vitro studies showed that the presence and sequence of the ScPif1 SM were critical for ATPase activity but not substrate binding.
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影响因子：
16
作者：
Ivessa, AS;Lenzmeier, BA;Zakian, VA
通讯作者：
Zakian, VA
影响因子：
14.9
作者：
Gorbalenya AE;Koonin EV;Donchenko AP;Blinov VM
通讯作者：
Blinov VM
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影响因子：
5.7
作者：
He, Xiaoping;Byrd, Alicia K.;Yun, Mi-Kyung;Pemble, Charles W.;Harrison, David;Yeruva, Laxmi;Dahl, Christopher;Kreuzer, Kenneth N.;Raney, Kevin D.;White, Stephen W.
通讯作者：
White, Stephen W.
影响因子：
3.8
作者：
Bochman ML;Sabouri N;Zakian VA
通讯作者：
Zakian VA
影响因子：
10.5
作者：
Azvolinsky, Anna;Dunaway, Stephen;Zakian, Virginia A.
通讯作者：
Zakian, Virginia A.