title: Polymerase manager protein UmuD directly regulates Escherichia coli DNA polymerase III α binding to ssDNA.
abstract: Replication by Escherichia coli DNA polymerase III is disrupted on encountering DNA damage. Consequently, specialized Y-family DNA polymerases are used to bypass DNA damage. The protein UmuD is extensively involved in modulating cellular responses to DNA damage and may play a role in DNA polymerase exchange for damage tolerance. In the absence of DNA, UmuD interacts with the α subunit of DNA polymerase III at two distinct binding sites, one of which is adjacent to the single-stranded DNA-binding site of α. Here, we use single molecule DNA stretching experiments to demonstrate that UmuD specifically inhibits binding of α to ssDNA. We predict using molecular modeling that UmuD residues D91 and G92 are involved in this interaction and demonstrate that mutation of these residues disrupts the interaction. Our results suggest that competition between UmuD and ssDNA for α binding is a new mechanism for polymerase exchange.
authors.affiliation: Department of Physics, Northeastern University, Boston, MA 02115, USA, Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, USA and Department of Chemical Sciences, Bridgewater State University, Bridgewater, MA 02325, USA.
authors.forename: Kathy R
authors.initials: KR
authors.lastname: Chaurasiya
authors.forename: Clarissa
authors.initials: C
authors.lastname: Ruslie
authors.forename: Michelle C
authors.initials: MC
authors.lastname: Silva
authors.forename: Lukas
authors.initials: L
authors.lastname: Voortman
authors.forename: Philip
authors.initials: P
authors.lastname: Nevin
authors.forename: Samer
authors.initials: S
authors.lastname: Lone
authors.forename: Penny J
authors.initials: PJ
authors.lastname: Beuning
authors.forename: Mark C
authors.initials: MC
authors.lastname: Williams
mesh_terms: D001667:Binding, Competitive; D004258:DNA Polymerase III / Q000737:chemistry / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009154:Mutation
chemical_list: D004277:DNA, Single-Stranded; D029968:Escherichia coli Proteins; C074904:DNA polymerase III, alpha subunit; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase; C055218:UmuD protein, E coli
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