Mcm10 self-association is mediated by an N-terminal coiled-coil domain.
Mcm10 self-association is mediated by an N-terminal coiled-coil domain.
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DOI：
10.1371/journal.pone.0070518
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Eichman BF
中科院分区：
文献类型：
--
作者：
Du W;Josephrajan A;Adhikary S;Bowles T;Bielinsky AK;Eichman BF
微小染色体维持蛋白10（Mcm 10）是一种重要的真核生物DNA结合复制因子，被认为是协调复制体内酶活性的支架。Mcm 10似乎作为低聚物而不是以其单体形式（或而不是作为单体）发挥作用。然而，已经发现各种直向同源物含有1、2、3、4或6个亚基，因此，这个问题仍然存在争议。在这里，我们表明，非洲爪蟾Mcm 10的自我关联介导的保守卷曲螺旋（CC）基序内的N-末端结构域（NTD）。在2.4 nm分辨率下对CC进行的晶体学分析显示了三螺旋束，这与溶液中二聚体和三聚体Mcm 10 CC的形成一致。突变的亚基界面的侧链破坏了CC和NTD的体外二聚化，如通过分析超离心监测的。此外，相同的突变也阻碍了全长蛋白质在体内的自我相互作用，如通过酵母双杂交测定所测量的。我们的结论是，Mcm 10可能形成二聚体或三聚体，以促进其在DNA复制过程中的多种功能。
Minichromosome maintenance protein 10 (Mcm10) is an essential eukaryotic DNA-binding replication factor thought to serve as a scaffold to coordinate enzymatic activities within the replisome. Mcm10 appears to function as an oligomer rather than in its monomeric form (or rather than as a monomer). However, various orthologs have been found to contain 1, 2, 3, 4, or 6 subunits and thus, this issue has remained controversial. Here, we show that self-association of Xenopus laevis Mcm10 is mediated by a conserved coiled-coil (CC) motif within the N-terminal domain (NTD). Crystallographic analysis of the CC at 2.4 Å resolution revealed a three-helix bundle, consistent with the formation of both dimeric and trimeric Mcm10 CCs in solution. Mutation of the side chains at the subunit interface disrupted in vitro dimerization of both the CC and the NTD as monitored by analytical ultracentrifugation. In addition, the same mutations also impeded self-interaction of the full-length protein in vivo, as measured by yeast-two hybrid assays. We conclude that Mcm10 likely forms dimers or trimers to promote its diverse functions during DNA replication.
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影响因子：
8
作者：
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通讯作者：
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影响因子：
14.9
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DOI：
10.1107/s0907444904019158
发表时间：
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影响因子：
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作者：
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DOI：
10.1073/pnas.1008502107
发表时间：
2010-11-16
影响因子：
11.1
作者：
Ciani, Barbara;Bjelic, Sasa;Kammerer, Richard A.
通讯作者：
Kammerer, Richard A.