Shell Scarlet S, Putnam Christopher D, Kolodner Richard D
Ludwig Institute for Cancer Research, University of California, San Diego, School of Medicine, 9500 Gilman Drive, La Jolla, CA 92093-0669, USA.
Mol Cell. 2007 May 25;26(4):565-78. doi: 10.1016/j.molcel.2007.04.024.
The eukaryotic MutS homolog complexes, Msh2-Msh6 and Msh2-Msh3, recognize mismatched bases in DNA during mismatch repair (MMR). The eukaryote-specific N-terminal regions (NTRs) of Msh6 and Msh3 have not been characterized other than by demonstrating that they contain an N-terminal PCNA-interacting motif. Here we have demonstrated genetically that the NTR of Msh6 has an important role in MMR that is partially redundant with PCNA binding. Small-angle X-ray scattering (SAXS) was used to determine the solution structure of the complex of PCNA with Msh2-Msh6 and with the isolated Msh6 NTR, revealing that the Msh6 NTR is a natively disordered domain that forms an extended tether between Msh6 and PCNA. Moreover, computational analysis of PCNA-interacting motifs in the S. cerevisiae proteome indicated that flexible linkers are a common theme for PCNA-interacting proteins that may serve to localize these binding partners without tightly restraining them to the immediate vicinity of PCNA.
真核生物错配修复蛋白MutS同源复合物Msh2-Msh6和Msh2-Msh3在错配修复(MMR)过程中识别DNA中的错配碱基。除了证明Msh6和Msh3的真核生物特异性N端区域(NTR)含有一个N端PCNA相互作用基序外,它们尚未得到表征。在这里,我们通过遗传学方法证明,Msh6的NTR在MMR中具有重要作用,且该作用与PCNA结合存在部分冗余。小角X射线散射(SAXS)用于确定PCNA与Msh2-Msh6以及分离的Msh6 NTR形成的复合物的溶液结构,结果表明Msh6 NTR是一个天然无序结构域,它在Msh6和PCNA之间形成了一个延伸的连接链。此外,对酿酒酵母蛋白质组中PCNA相互作用基序的计算分析表明,柔性连接子是PCNA相互作用蛋白的一个共同特征,这可能有助于定位这些结合伴侣,而不会将它们紧密限制在PCNA的紧邻区域。