PMID:15210708

Citation	Bertram, JG, Bloom, LB, O'Donnell, M and Goodman, MF (2004) Increased dNTP binding affinity reveals a nonprocessive role for Escherichia coli beta clamp with DNA polymerase IV. J. Biol. Chem. 279:33047-50
Abstract	Replication forks often stall at undamaged or damaged template sites in Escherichia coli. Subsequent resumption of DNA synthesis occurs by replacing DNA polymerase III, which is bound to DNA by the beta-sliding clamp, with one of three damage-induced DNA polymerases II, IV, or V. The principal role of the beta clamp is to tether the normally weakly bound polmerases to DNA thereby increasing their processivities. DNA polymerase IV binds dNTP substrates with about 10-fold lower affinity compared with the other E. coli polymerases, which if left unchecked could hinder its ability to synthesize DNA in vivo. Here we report a new property for the beta clamp, which when bound to DNA polymerase IV results in a large increase in dNTP binding affinity that concomitantly increases the efficiency of nucleotide incorporation at normal and transiently slipped mispaired primer/template ends. Primer-template DNA slippage resulting in single nucleotide deletions is a biological hallmark of DNA polymerase IV infidelity responsible for enhancing cell fitness in response to stress. We show that the increased DNA polymerase IV-dNTP binding affinity is an intrinsic property of the DNA polymerase IV-beta clamp interaction and not an indirect consequence of an increased binding of DNA polymerase IV to DNA.
Links	PubMed Online version:10.1074/jbc.C400265200
Keywords	Base Pair Mismatch; Binding Sites; DNA/chemistry; DNA/genetics; DNA/metabolism; DNA Polymerase III/metabolism; DNA Polymerase beta/chemistry; DNA Polymerase beta/metabolism; DNA Primers/metabolism; Deoxyguanine Nucleotides/metabolism; Deoxyribonucleotides/chemistry; Deoxyribonucleotides/metabolism; Escherichia coli/enzymology; Fluorescence Polarization; Frameshift Mutation; Kinetics; Oligodeoxyribonucleotides/chemistry; Oligodeoxyribonucleotides/metabolism; Templates, Genetic; Thymine Nucleotides/metabolism
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