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Mismatch Recognition by Saccharomyces cerevisiae Msh2-Msh6: Role of Structure and Dynamics

Molecular Biology and Biochemistry Department, Molecular Biophysics Program, Wesleyan University, Middletown, CT 06459, USA
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Int. J. Mol. Sci. 2019, 20(17), 4271; https://doi.org/10.3390/ijms20174271
Received: 7 August 2019 / Revised: 27 August 2019 / Accepted: 27 August 2019 / Published: 31 August 2019
(This article belongs to the Section Physical Chemistry and Chemical Physics)
The mismatch repair (MMR) pathway maintains genome integrity by correcting errors such as mismatched base pairs formed during DNA replication. In MMR, Msh2–Msh6, a heterodimeric protein, targets single base mismatches and small insertion/deletion loops for repair. By incorporating the fluorescent nucleoside base analog 6-methylisoxanthopterin (6-MI) at or adjacent to a mismatch site to probe the structural and dynamic elements of the mismatch, we address how Msh2–Msh6 recognizes these mismatches for repair within the context of matched DNA. Fluorescence quantum yield and rotational correlation time measurements indicate that local base dynamics linearly correlate with Saccharomyces cerevisiae Msh2–Msh6 binding affinity where the protein exhibits a higher affinity (KD ≤ 25 nM) for mismatches that have a significant amount of dynamic motion. Energy transfer measurements measuring global DNA bending find that mismatches that are both well and poorly recognized by Msh2–Msh6 experience the same amount of protein-induced bending. Finally, base-specific dynamics coupled with protein-induced blue shifts in peak emission strongly support the crystallographic model of directional binding, in which Phe 432 of Msh6 intercalates 3′ of the mismatch. These results imply an important role for local base dynamics in the initial recognition step of MMR. View Full-Text
Keywords: Msh2–Msh6; MutSα; DNA repair; fluorescence; Förster resonance energy transfer (FRET); 6-methylisoxanthopterin (6-MI); protein-DNA interactions; induced fit; base dynamics; mismatch repair (MMR) Msh2–Msh6; MutSα; DNA repair; fluorescence; Förster resonance energy transfer (FRET); 6-methylisoxanthopterin (6-MI); protein-DNA interactions; induced fit; base dynamics; mismatch repair (MMR)
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Li, Y.; Lombardo, Z.; Joshi, M.; Hingorani, M.M.; Mukerji, I. Mismatch Recognition by Saccharomyces cerevisiae Msh2-Msh6: Role of Structure and Dynamics. Int. J. Mol. Sci. 2019, 20, 4271.

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