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

Kouta Mayanagi, Shinichi Kiyonari, Mihoko Saito, Tsuyoshi Shirai, Yoshizumi Ishino, Kosuke Morikawa

Research output: Contribution to journalArticle

50 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)4647-4652
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number12
DOIs
Publication statusPublished - Mar 24 2009

All Science Journal Classification (ASJC) codes

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