title: UmuDAb: An Error-Prone Polymerase Accessory Homolog Whose N-Terminal Domain Is Required for Repression of DNA Damage Inducible Gene Expression in Acinetobacter baylyi.
abstract: In many bacteria, the DNA damage response induces genes (SOS genes) that were repressed by LexA. LexA represses transcription by binding to SOS promoters via a helix-turn-helix motif in its N-terminal domain (NTD). Upon DNA damage, LexA cleaves itself and allows induction of transcription. In Acinetobacter baumannii and Acinetobacter baylyi, multiple genes are induced by DNA damage, and although the Acinetobacter genus lacks LexA, a homolog of the error-prone polymerase subunit UmuD, called UmuDAb, regulates some DNA damage-induced genes. The mechanism of UmuDAb regulation has not been determined. We constructed UmuDAb mutant strains of A. baylyi to test whether UmuDAb mediates gene regulation through LexA-like repressor actions consisting of relief of repression through self-cleavage after DNA damage. Real-time quantitative PCR experiments in both a null umuDAb mutant and an NTD mutant showed that the DNA damage-inducible, UmuDAb-regulated gene ddrR was highly expressed even in the absence of DNA damage. Protein modeling identified a potential LexA-like helix-turn-helix structure in the UmuDAb NTD, which when disrupted, also relieved ddrR and umuDAb repression under non-inducing conditions. Mutations in a putative SOS box in the shared umuDAb-ddrR promoter region similarly relieved these genes' repression under non-inducing conditions. Conversely, cells possessing a cleavage-deficient UmuDAb were unable to induce gene expression after MMC-mediated DNA damage. This evidence of a UmuDAb repressor mechanism was contrasted with the failure of umuDAb to complement an Escherichia coli umuD mutant for UmuD error-prone DNA replication activity. Similarly, A. baumannii null umuDAb mutant cells did not have a reduced UmuD'2UmuC-mediated mutation rate after DNA damage, suggesting that although this UmuDAb protein may have evolved from a umuDC operon in this genus, it now performs a LexA-like repressor function for a sub-set of DNA damage-induced genes.
authors.affiliation: Department of Biology and Chemistry, Morehead State University, Morehead, KY, United States of America.
authors.forename: Travis A
authors.initials: TA
authors.lastname: Witkowski
authors.affiliation: Department of Biology and Chemistry, Morehead State University, Morehead, KY, United States of America.
authors.forename: Alison N
authors.initials: AN
authors.lastname: Grice
authors.affiliation: Department of Biology and Chemistry, Morehead State University, Morehead, KY, United States of America.
authors.forename: DeAnna B
authors.initials: DB
authors.lastname: Stinnett
authors.affiliation: Department of Biology and Chemistry, Morehead State University, Morehead, KY, United States of America.
authors.forename: Whitney K
authors.initials: WK
authors.lastname: Wells
authors.affiliation: Department of Biology and Chemistry, Morehead State University, Morehead, KY, United States of America.
authors.forename: Megan A
authors.initials: MA
authors.lastname: Peterson
authors.affiliation: Department of Biology and Chemistry, Morehead State University, Morehead, KY, United States of America.
authors.forename: Janelle M
authors.initials: JM
authors.lastname: Hare
mesh_terms: D000150:Acinetobacter / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D004249:DNA Damage / Q000235:genetics*; D004261:DNA Replication / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D012097:Repressor Proteins / Q000235:genetics*; D013014:SOS Response, Genetics; D012697:Serine Endopeptidases / Q000235:genetics; D000099160:LexA Repressor Protein
chemical_list: D001426:Bacterial Proteins; D004259:DNA-Directed DNA Polymerase; D029968:Escherichia coli Proteins; D012097:Repressor Proteins; D012697:Serine Endopeptidases; D000099160:LexA Repressor Protein; C055218:UmuD protein, E coli