文章基本信息

标题：Mechanism of replication machinery assembly as revealed by the DNA ligase–PCNA–DNA complex architecture
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作者：Kouta Mayanagi ; Shinichi Kiyonari ; Mihoko Saito 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2009
卷号：106
期号：12
页码：4647-4652
DOI：10.1073/pnas.0811196106
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：The 3D structure of the ternary complex, consisting of DNA ligase, the proliferating cell nuclear antigen (PCNA) clamp, and DNA, was investigated by single-particle analysis. This report presents the structural view, where the crescent-shaped DNA ligase with 3 distinct domains surrounds the central DNA duplex, encircled by the closed PCNA ring, thus forming a double-layer structure with dual contacts between the 2 proteins. The relative orientations of the DNA ligase domains, which remarkably differ from those of the known crystal structures, suggest that a large domain rearrangement occurs upon ternary complex formation. A second contact was found between the PCNA ring and the middle adenylation domain of the DNA ligase. Notably, the map revealed a substantial DNA tilt from the PCNA ring axis. This structure allows us to propose a switching mechanism for the replication factors operating on the PCNA ring.
关键词：DNA replication ; electron microscopy ; single-particle analysis ; DNA sliding clamp ; protein-DNA complex