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Mechanism of replication machinery assembly as revealed by the DNA ligase-PCNA-DNA complex architecture.


ABSTRACT: 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.

SUBMITTER: Mayanagi K 

PROVIDER: S-EPMC2660782 | biostudies-literature | 2009 Mar

REPOSITORIES: biostudies-literature

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Mechanism of replication machinery assembly as revealed by the DNA ligase-PCNA-DNA complex architecture.

Mayanagi Kouta K   Kiyonari Shinichi S   Saito Mihoko M   Shirai Tsuyoshi T   Ishino Yoshizumi Y   Morikawa Kosuke K  

Proceedings of the National Academy of Sciences of the United States of America 20090302 12


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 di  ...[more]

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