title: Roles for the transcription elongation factor NusA in both DNA repair and damage tolerance pathways in Escherichia coli.
abstract: We report observations suggesting that the transcription elongation factor NusA promotes a previously unrecognized class of transcription-coupled repair (TCR) in addition to its previously proposed role in recruiting translesion synthesis (TLS) DNA polymerases to gaps encountered during transcription. Earlier, we reported that NusA physically and genetically interacts with the TLS DNA polymerase DinB (DNA pol IV). We find that Escherichia coli nusA11(ts) mutant strains, at the permissive temperature, are highly sensitive to nitrofurazone (NFZ) and 4-nitroquinolone-1-oxide but not to UV radiation. Gene expression profiling suggests that this sensitivity is unlikely to be due to an indirect effect on gene expression affecting a known DNA repair or damage tolerance pathway. We demonstrate that an N(2)-furfuryl-dG (N(2)-f-dG) lesion, a structural analog of the principal lesion generated by NFZ, blocks transcription by E. coli RNA polymerase (RNAP) when present in the transcribed strand, but not when present in the nontranscribed strand. Our genetic analysis suggests that NusA participates in a nucleotide excision repair (NER)-dependent process to promote NFZ resistance. We provide evidence that transcription plays a role in the repair of NFZ-induced lesions through the isolation of RNAP mutants that display altered ability to survive NFZ exposure. We propose that NusA participates in an alternative class of TCR involved in the identification and removal of a class of lesion, such as the N(2)-f-dG lesion, which are accurately and efficiently bypassed by DinB in addition to recruiting DinB for TLS at gaps encountered by RNAP.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Susan E
authors.initials: SE
authors.lastname: Cohen
authors.forename: Cindi A
authors.initials: CA
authors.lastname: Lewis
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Mooney
authors.forename: Michael A
authors.initials: MA
authors.lastname: Kohanski
authors.forename: James J
authors.initials: JJ
authors.lastname: Collins
authors.forename: Robert
authors.initials: R
authors.lastname: Landick
authors.forename: Graham C
authors.initials: GC
authors.lastname: Walker
mesh_terms: D015112:4-Nitroquinoline-1-oxide / Q000494:pharmacology; D000890:Anti-Infective Agents / Q000494:pharmacology; D026481:Blotting, Far-Western; D004249:DNA Damage; D004260:DNA Repair; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D050296:Microbial Viability / Q000187:drug effects; D008856:Microscopy, Fluorescence; D009154:Mutation; D009583:Nitrofurazone / Q000494:pharmacology; D010445:Peptide Elongation Factors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015363:Quinolones / Q000494:pharmacology; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013696:Temperature; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic / Q000187:drug effects; D035602:Transcriptional Elongation Factors
chemical_list: C120195:4-nitroquinolone-1-oxide; D000890:Anti-Infective Agents; D029968:Escherichia coli Proteins; D010445:Peptide Elongation Factors; D015363:Quinolones; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; C068727:nusA protein, E coli; D049452:Green Fluorescent Proteins; D015112:4-Nitroquinoline-1-oxide; D011938:Rec A Recombinases; D012321:DNA-Directed RNA Polymerases; C113275:RNA polymerase alpha subunit; D009583:Nitrofurazone