Structural determinant for switching between the polymerase and exonuclease modes in the PCNA-replicative DNA polymerase complex

Hirokazu Nishida, Kouta Mayanagi, Shinichi Kiyonari, Yuichi Sato, Takuji Oyama, Yoshizumi Ishino, Kosuke Morikawa

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Proliferating cell nuclear antigen (PCNA) is responsible for the processivity of DNA polymerase. We determined the crystal structure of Pyrococcus furiosus DNA polymerase (PfuPol) complexed with the cognate monomeric PCNA, which allowed us to construct a convincing model of the polymerase-PCNA ring interaction, with unprecedented configurations of the two molecules. Electron microscopic analyses indicated that this complex structure exists in solution. Our structural study revealed that an interaction occurs between a stretched loop of PCNA and the PfuPol Thumb domain, in addition to the authentic PCNA-polymerase recognition site (PIP box). Comparisons of the present structure with the previously reported structures of polymerases complexed with DNA, suggested that the second interaction plays a crucial role in switching between the polymerase and exonuclease modes, by inducing a PCNA-polymerase complex configuration that favors synthesis over editing. This putative mechanism for fidelity control of replicative DNA polymerases is supported by experiments, in which mutations at the second interaction site caused enhancements in the exonuclease activity in the presence of PCNA.

Original languageEnglish
Pages (from-to)20693-20698
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number49
DOIs
Publication statusPublished - Dec 8 2009

All Science Journal Classification (ASJC) codes

  • General

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