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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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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Supplementary Fig. 1
CAG-hairpin binding changes stoichiometry, not composition, of protein/DNA complexes. (PDF 98 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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DOI: https://doi.org/10.1038/nsmb965
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