title: Helicobacter pylori FlgR is an enhancer-independent activator of sigma54-RNA polymerase holoenzyme.
abstract: Helicobacter pylori FlgR activates transcription with sigma54-RNA polymerase holoenzyme (sigma54-holoenzyme) from at least five flagellar operons. Activators of sigma54-holoenzyme generally bind enhancer sequences located >70 bp upstream of the promoter and contact sigma54-holoenzyme bound at the promoter through DNA looping to activate transcription. H. pylori FlgR lacks the carboxy-terminal DNA-binding domain present in most sigma54-dependent activators. As little as 42 bp of DNA upstream of the flaB promoter and 26 bp of DNA sequence downstream of the transcriptional start site were sufficient for efficient FlgR-mediated expression from a flaB'-'xylE reporter gene in H. pylori, indicating that FlgR does not use an enhancer to activate transcription. Other examples of sigma54-dependent activators that lack a DNA-binding domain include Chlamydia trachomatis CtcC and activators from the other Chlamydia spp. whose genomes have been sequenced. FlgR from Helicobacter hepaticus and Campylobacter jejuni, which are closely related to H. pylori, appear to have carboxy-terminal DNA-binding domains, suggesting that the loss of the DNA-binding domain from H. pylori FlgR occurred after the divergence of these bacterial species. Removal of the amino-terminal regulatory domain of FlgR resulted in a constitutively active form of the protein that activated transcription from sigma54-dependent genes in Escherichia coli. The truncated FlgR protein also activated transcription with E. coli sigma54-holoenzyme in an in vitro transcription assay.
authors.affiliation: Department of Microbiology, University of Georgia, Athens, GA 30602, USA.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Brahmachary
authors.forename: Mona G
authors.initials: MG
authors.lastname: Dashti
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Olson
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Hoover
mesh_terms: D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D016123:Campylobacter jejuni / Q000235:genetics; D002692:Chlamydia trachomatis / Q000235:genetics; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D005408:Flagellin / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D044923:Helicobacter hepaticus / Q000235:genetics; D016480:Helicobacter pylori / Q000235:genetics* / Q000378:metabolism*; D015151:Immunoblotting; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D051877:RNA Polymerase Sigma 54; D012808:Sigma Factor / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D012808:Sigma Factor; C497751:rpoN protein, E coli; D005408:Flagellin; C416532:flaB flagellin; D012321:DNA-Directed RNA Polymerases; D051877:RNA Polymerase Sigma 54; D001616:beta-Galactosidase