Characterization of the interactome of the human MutL homologues MLH1, PMS1, and PMS2
Characterization of the interactome of the human MutL homologues MLH1, PMS1, and PMS2
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DOI：
10.1074/jbc.m609989200
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发表时间：
2007-02-02
影响因子：
4.8
通讯作者：
Jiricny, Josef
中科院分区：
文献类型：
--
作者：
Cannavo, Elda;Gerrits, Bertran;Jiricny, Josef
复制后错配修复（MMR）涉及至少20种多肽的协同作用。虽然最小的人类MMR系统最近已在体外重建，从不同的真核生物的遗传证据表明，MMR过程的一些步骤可能是由一个以上的蛋白质。此外，MMR蛋白也参与DNA代谢的其他途径，但它们在这些过程中的确切作用尚不清楚。在试图获得新的见解MMR蛋白在人类细胞中的功能，我们寻找相互作用的合作伙伴的MutL同源MLH1和PMS2的串联亲和纯化和PMS1的大规模免疫沉淀。除了已知在MMR过程中与MutL同源物相互作用的蛋白质外，质谱分析还鉴定了许多其他多肽，其中一些多肽以非常高的亲和力与上述蛋白质结合。尽管这些相互作用物中的一些可能代表错配修复体的新成员，但其他相互作用物似乎涉及MutL同源物在生物过程中的作用，所述生物过程从细胞内转运通过细胞信号传导到细胞形态、重组和泛素化。
Postreplicative mismatch repair (MMR) involves the concerted action of at least 20 polypeptides. Although the minimal human MMR system has recently been reconstituted in vitro, genetic evidence from different eukaryotic organisms suggests that some steps of the MMR process may be carried out by more than one protein. Moreover, MMR proteins are involved also in other pathways of DNA metabolism, but their exact role in these processes is unknown. In an attempt to gain novel insights into the function of MMR proteins in human cells, we searched for interacting partners of the MutL homologues MLH1 and PMS2 by tandem affinity purification and of PMS1 by large scale immunoprecipitation. In addition to proteins known to interact with the MutL homologues during MMR, mass spectrometric analyses identified a number of other polypeptides, some of which bound to the above proteins with very high affinity. Whereas some of these interactors may represent novel members of the mismatch repairosome, others appear to implicate the MutL homologues in biological processes ranging from intracellular transport through cell signaling to cell morphology, recombination, and ubiquitylation.