title: Antagonistic regulation of Escherichia coli ribosomal RNA rrnB P1 promoter activity by GreA and DksA.
abstract: The Escherichia coli proteins DksA, GreA, and GreB are all structural homologs that bind the secondary channel of RNA polymerase (RNAP) but are thought to act at different levels of transcription. DksA, with its co-factor ppGpp, inhibits rrnB P1 transcription initiation, whereas GreA and GreB activate RNAP to cleave back-tracked RNA during elongational pausing. Here, in vivo and in vitro evidence reveals antagonistic regulation of rrnB P1 transcription initiation by Gre factors (particularly GreA) and DksA; GreA activates and DksA inhibits. DksA inhibition is epistatic to GreA activation. Both modes of regulation are ppGpp-independent in vivo but DksA inhibition requires ppGpp in vitro. Kinetic experiments and studies of rrnB P1-RNA polymerase complexes suggest that GreA mediates conformational changes at an initiation step in the absence of NTP substrates, even before DksA acts. GreA effects on rrnB P1 open complex conformation reveal a new feature of GreA distinct from its general function in elongation. Our findings support the idea that a balance of the interactions between the three secondary channel-binding proteins and RNAP can provide a new mode for regulating transcription.
authors.affiliation: Laboratory of Molecular Genetics, NICHD, National Institutes of Health, Bethesda, Maryland 20892-2785, USA.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Potrykus
authors.forename: Daniel
authors.initials: D
authors.lastname: Vinella
authors.forename: Helen
authors.initials: H
authors.lastname: Murphy
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Szalewska-Palasz
authors.forename: Richard
authors.initials: R
authors.lastname: D'Ari
authors.forename: Michael
authors.initials: M
authors.lastname: Cashel
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000378:metabolism; D004843:Epistasis, Genetic; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008954:Models, Biological; D046912:Multiprotein Complexes; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000235:genetics* / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D012407:rRNA Operon
chemical_list: D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C079668:GreA protein, E coli; D046912:Multiprotein Complexes; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D014157:Transcription Factors; C090751:dksA protein, E coli; D012321:DNA-Directed RNA Polymerases