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(CAG)n-hairpin DNA binds to Msh2–Msh3 and changes properties of mismatch recognition

Nature Structural & Molecular Biology volume 12, pages 663–670 (2005)Cite this article

A Corrigendum to this article was published on 01 September 2005

Abstract

Cells have evolved sophisticated DNA repair systems to correct damaged DNA. However, the human DNA mismatch repair protein Msh2–Msh3 is involved in the process of trinucleotide (CNG) DNA expansion rather than repair. Using purified protein and synthetic DNA substrates, we show that Msh2–Msh3 binds to CAG-hairpin DNA, a prime candidate for an expansion intermediate. CAG-hairpin binding inhibits the ATPase activity of Msh2–Msh3 and alters both nucleotide (ADP and ATP) affinity and binding interfaces between protein and DNA. These changes in Msh2–Msh3 function depend on the presence of A·A mispaired bases in the stem of the hairpin and on the hairpin DNA structure per se. These studies identify critical functional defects in the Msh2–Msh3–CAG hairpin complex that could misdirect the DNA repair process.

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Figure 1: Msh2–Msh3 causes CAG expansion in a coding sequence.
Figure 2: Msh2–Msh3 binds to CAG-hairpin DNA with high affinity.
Figure 4: Binding of Msh2–Msh3 to CAG hairpins inhibits ATP hydrolysis.
Figure 3: Msh2–Msh3 binding to the CAG-hairpin is greater than 1:1.
Figure 5: Multiple A•A mismatches inhibit ATPase activity of Msh2–Msh3.
Figure 6: Relative affinity of Msh2–Msh3 for nucleotide is DNA dependent.
Figure 7: Msh2–Msh3 binds differently to DNA loops and hairpins.
Figure 8: Duplex length does not affect ATPase activity of Msh2–Msh3 when bound to hairpin DNA.
Figure 9: Uncoupling models for Msh2–Msh3 bound to CAG hairpins.

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Acknowledgements

We thank J.P. Carney for discussions and D.O. Toft for critical reading of the manuscript. This work was funded by the US National Institutes of Health. This work was funded by NIH GM-066359 and NINDS-R01NS40738 (C.T.M.); NIH CA-0955690 (T.M.W.); NIH-GM52426 (J.J.H.); and NIH-CA84301 and NIH-ES11040 (R.K.).

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Authors and Affiliations

  1. Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic Rochester, Rochester, 55905, Minnesota, USA

    Barbara A L Owen, Maoyi Lai, John D Badger III & Cynthia T McMurray

  2. Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, 14642, New York, USA

    Zungyoon Yang & Jeffrey J Hayes

  3. Department of Radiation Oncology, University of Maryland, Baltimore, 21201, Maryland, USA

    Maciez Gajek & Teresa M Wilson

  4. Department of Cell Biology, Albert Einstein College, Bronx, 10461, New York, USA

    Winfried Edelmann

  5. Harvard Medical School-Partners Healthcare for Genetics and Genomics, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Raju Kucherlapati

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Supplementary information

Supplementary Fig. 1

CAG-hairpin binding changes stoichiometry, not composition, of protein/DNA complexes. (PDF 98 kb)

Supplementary Methods (PDF 117 kb)

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Owen, B., Yang, Z., Lai, M. et al. (CAG)n-hairpin DNA binds to Msh2–Msh3 and changes properties of mismatch recognition. Nat Struct Mol Biol 12, 663–670 (2005). https://doi.org/10.1038/nsmb965

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