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PubMed · 16546997

The beta sliding clamp binds to multiple sites within MutL and MutS.

Francisco J López de Saro·Martin G Marinus·Paul Modrich·Mike O'Donnell
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Abstract

The MutL and MutS proteins are the central components of the DNA repair machinery that corrects mismatches generated by DNA polymerases during synthesis. We find that MutL interacts directly with the beta sliding clamp, a ring-shaped dimeric protein that confers processivity to DNA polymerases by tethering them to their substrates. Interestingly, the interaction of MutL with beta only occurs in the presence of single-stranded DNA. We find that the interaction occurs via a loop in MutL near the ATP-binding site. The binding site of MutL on beta locates to the hydrophobic pocket between domains two and three of the clamp. Site-specific replacement of two residues in MutL diminished interaction with beta without disrupting MutL function with helicase II. In vivo studies reveal that this mutant MutL is no longer functional in mismatch repair. In addition, the human MLH1 has a close match to the proliferating cell nuclear antigen clamp binding motif in the region that corresponds to the beta interaction site in Escherichia coli MutL, and a peptide corresponding to this site binds proliferating cell nuclear antigen. The current report also examines in detail the interaction of beta with MutS. We find that two distinct regions of MutS interact with beta. One is located near the C terminus and the other is close to the N terminus, within the mismatch binding domain. Complementation studies using genes encoding different MutS mutants reveal that the N-terminal beta interaction motif on MutS is essential for activity in vivo, but the C-terminal interaction site for beta is not. In light of these results, we propose roles for the beta clamp in orchestrating the sequence of events that lead to mismatch repair in the cell.

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Adaptor Proteins, Signal TransducingAdenosine TriphosphatasesAmino Acid SequenceCarrier ProteinsDNA RepairDNA, Single-StrandedDose-Response Relationship, DrugEscherichia coliEscherichia coli ProteinsGenetic Complementation TestHumansModels, MolecularMolecular Sequence DataMutL Protein Homolog 1MutL ProteinsMutS DNA Mismatch-Binding ProteinNuclear ProteinsProtein BindingSequence Homology, Amino AcidJournal ArticleResearch Support, N.I.H., Extramural

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Francisco J López de Saro, Martin G Marinus, Paul Modrich, Mike O'Donnell. 2006-03-16. The beta sliding clamp binds to multiple sites within MutL and MutS.. https://doi.org/10.1074/jbc.m601264200

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