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The nucleotide binding dynamics of human MSH2–MSH3 are lesion dependent

Nature Structural & Molecular Biology volume 16, pages 550–557 (2009)Cite this article

An Erratum to this article was published on 01 August 2009

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Abstract

Here we report that the human DNA mismatch complex MSH2–MSH3 recognizes small loops by a mechanism different from that of MSH2–MSH6 for single-base mismatches. The subunits MSH2 and MSH3 can bind either ADP or ATP with similar affinities. Upon binding to a DNA loop, however, MSH2–MSH3 adopts a single 'nucleotide signature', in which the MSH2 subunit is occupied by an ADP molecule and the MSH3 subunit is empty. Subsequent ATP binding and hydrolysis in the MSH3 subunit promote ADP-ATP exchange in the MSH2 subunit to yield a hydrolysis-independent ATP-MSH2–MSH3-ADP intermediate. Human MSH2–MSH3 and yeast Msh2–Msh6 both undergo ADP-ATP exchange in the Msh2 subunit but, apparently, have opposite requirements for ATP hydrolysis: ADP release from DNA-bound Msh2–Msh6 requires ATP stabilization in the Msh6 subunit, whereas ADP release from DNA-bound MSH2–MSH3 requires ATP hydrolysis in the MSH3 subunit. We propose a model in which lesion binding converts MSH2–MSH3 into a distinct nucleotide-bound form that is poised to be a molecular sensor for lesion specificity.

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Figure 1: Human MSH2–MSH6 binds ADP and ATP in a subunit-specific manner.
Figure 2: Human MSH2–MSH3 binds ATP and ADP with equal apparent affinity in both subunits.
Figure 3: Binding of either ADP or ATP to one subunit of MSH2–MSH3 excludes binding in the other.
Figure 4: ADP-bound forms of MSH2–MSH3 are stable in solution.
Figure 5: ADP-MSH2–MSH3-empty stably binds DNA.
Figure 6: ATP hydrolysis enhances ADP-ATP exchange in the MSH2 subunit of (CA)4-loop bound MSH2–MSH3, and promotes a hydrolysis-independent state.
Figure 7: Model for loop recognition by MSH2–MSH3.

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Change history

  • 18 May 2009

    In the version of this article initially published, the sentence referring to Figure 2c (page 3, first paragraph) was incorrect. It should read: “To further test the stochastic binding of nucleotides, we simultaneously added a constant amount (5 µM) of labeled ADP(+Mg2+) and increasing concentrations of unlabeled AMP-PNP(+Mg2+) to MSH2–MSH3 (Fig. 2c, left).” The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank R. Weinshilboum, T.C. Wood and L.J. Maher III, for providing access to crucial equipment and I. Kovtun and J. Trushina for helpful comments. This work was supported by the Mayo Foundation and the US National Institutes of Health grants NS40738 (C.T.M.), GM066359 (C.T.M.) and CA092584 (C.T.M.).

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, Minnesota, USA

    Barbara A L Owen & Cynthia T McMurray

  2. Department of Internal Medicine, Mayo Clinic College of Medicine, Rochester, Minnesota, USA

    Barbara A L Owen

  3. Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA

    Walter H Lang & Cynthia T McMurray

  4. Department of Pharmacology and Experimental Therapeutics, Mayo Clinic College of Medicine, Rochester, Minnesota, USA

    Cynthia T McMurray

  5. Neuroscience Program, Mayo Clinic College of Medicine, Rochester, Minnesota, USA

    Cynthia T McMurray

Authors
  1. Barbara A L Owen
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  2. Walter H Lang
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Contributions

B.A.L.O. conceived and performed all of the nucleotide binding experiments under nonhydrolyzing conditions, determined the nucleotide binding stoichiometry to MSH2–MSH3 and wrote a substantial portion of the manuscript; W.H.L. conceived, performed and wrote text for the nucleotide binding experiments under hydrolyzing conditions. C.T.M. is responsible for the overall experimental design and analysis, and participated in the writing of the manuscript.

Corresponding author

Correspondence to Cynthia T McMurray.

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Supplementary Figures 1–7, Supplementary Table 1 and Supplementary Methods (PDF 1475 kb)

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Owen, B., H Lang, W. & McMurray, C. The nucleotide binding dynamics of human MSH2–MSH3 are lesion dependent. Nat Struct Mol Biol 16, 550–557 (2009). https://doi.org/10.1038/nsmb.1596

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