Single-molecule trajectory analysis has suggested DNA repair proteins may carry out a one-dimensional (1D) search on naked DNA encompassing >10,000 nucleotides. Organized cellular DNA (chromatin) presents substantial barriers to such lengthy searches. Using dynamic single-molecule fluorescence resonance energy transfer, we determined that the mismatch repair (MMR) initiation protein MutS forms a transient clamp that scans duplex DNA for mismatched nucleotides by 1D diffusion for 1 s (~700 base pairs) while in continuous rotational contact with the DNA. Mismatch identification provokes ATP binding (3 s) that induces distinctly different MutS sliding clamps with unusual stability on DNA (~600 s), which may be released by adjacent single-stranded DNA (ssDNA). These observations suggest that ATP transforms short-lived MutS lesion scanning clamps into highly stable MMR signaling clamps that are capable of competing with chromatin and recruiting MMR machinery, yet are recycled by ssDNA excision tracts.
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We thank J.-H. Park for helping with the FRET experiments. This work was supported by National Research Foundation of Korea grants funded by the Korean government (MEST) (no. 2008-0061211, no. 2009-351-C00118; C.J. postdoctoral fellowship), the Brain Korea 21 project, a POSTECH Basic Science Research Institute Grant (J.-B.L.), HRF-2008-314-C00218 (C.B.) and US National Institutes of Health grant CA067007 (R.F.).
The authors declare no competing financial interests.
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Jeong, C., Cho, WK., Song, KM. et al. MutS switches between two fundamentally distinct clamps during mismatch repair. Nat Struct Mol Biol 18, 379–385 (2011). https://doi.org/10.1038/nsmb.2009
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