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Proliferating cell nuclear antigen and Msh2p-Msh6p interact to form an active mispair recognition complex

Nature Genetics volume 26, pages 375–378 (2000)Cite this article

Abstract

Proliferating cell nuclear antigen (PCNA) is required for mismatch repair (MMR) and has been shown to interact with complexes containing Msh2p or MLH1 (refs 1–4). PCNA has been implicated to act in MMR before and during the DNA synthesis step, although the biochemical basis for the role of PCNA early in MMR is unclear1,3,5. Here we observe an interaction between PCNA and Msh2p-Msh6p mediated by a specific PCNA-binding site present in Msh6p. An msh6 mutation that eliminated the PCNA-binding site caused a mutator phenotype and a defect in the interaction with PCNA. The association of PCNA with Msh2p-Msh6p stimulated the preferential binding of Msh2p-Msh6p to DNA containing mispaired bases. Mutant PCNA proteins encoded by MMR-defective pol30 alleles were defective for interaction with Msh2p-Msh6p and for stimulation of mispair binding by Msh2p-Msh6p. Our results suggest that PCNA functions directly in mispair recognition and that mispair recognition requires a higher-order complex containing proteins in addition to Msh2p-Msh6p.

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Figure 1: Identification of a PCNA-binding site in MMR proteins.
Figure 2: Physical interaction between Msh2p-Msh6p and PCNA.
Figure 3: Effect of PCNA on Msh2p-Msh6p binding to mispairs.

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Acknowledgements

We thank N.S. Amin, R. Das Gupta, P. Lau, T. Nakagawa and K. Schmidt for helpful discussions and comments on the manuscript, and J. Weger and J. Green for DNA sequencing. This work was supported by National Institutes of Health grant GM50006 to R.D.K. and a fellowship from the Jane Coffin Childs Memorial Fund to H.F.-R.

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  1. Cancer Center and Department of Medicine, Ludwig Institute for Cancer Research, University of California-San Diego School of Medicine, La Jolla, California, USA

    Hernan Flores-Rozas, Delbert Clark & Richard D. Kolodner

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Correspondence to Richard D. Kolodner.

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Flores-Rozas, H., Clark, D. & Kolodner, R. Proliferating cell nuclear antigen and Msh2p-Msh6p interact to form an active mispair recognition complex. Nat Genet 26, 375–378 (2000). https://doi.org/10.1038/81708

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