Growth phase-dependent regulation of target gene promoters for binding of the essential orphan response regulator HP1043 of Helicobacter pylori.
Helicobacter pylori encodes three two-component systems and two orphan response regulators (RRs) that are predicted to be involved in transcriptional regulation. The HP1043 gene encodes an essential OmpR-like RR, 1043RR, for which no histidine kinase has been identified. Gel filtration and cross-linking experiments on the purified 1043RR protein reveals that this protein is a dimer and in vivo dimerization assays localize the dimerization to the N-terminal regulatory domain. DNA-binding studies have revealed two targets for specific binding of the 1043RR protein and moreover, phosphorylation of the protein was not needed for the activation of binding. Footprinting analysis demonstrated that the 1043RR protein binds to its own promoter, P(1043), overlapping the -35 promoter element from positions -17 to -45, suggesting that this protein is autoregulatory. In addition, it binds at a similar location, spanning nucleotides from positions -22 to -51 at the promoter of the methyl-accepting chemotaxis tlpB gene, P(tlpB). A possible inverted repeat was identified in the binding sites of both promoters. In an attempt to overexpress 1043RR in H. pylori, the 10-fold induction in transcription of a second copy of HP1043 with use of an inducible promoter failed to increase cellular levels of the RR protein, suggesting that 1043RR is tightly regulated at a posttranscriptional level. The P(1043) and P(tlpB) promoters were demonstrated to be coordinately regulated in response to growth phase in H. pylori. The essential role of HP1043 in encoding a cell cycle regulator is discussed.
Delany
Isabel
I
Department of Molecular Biology, IRIS, Chiron SpA, Via Fiorentina 1, 53100 Siena, Italy.
Spohn
Gunther
G
Rappuoli
Rino
R
Scarlato
Vincenzo
V
D001426:Bacterial Proteins; D002453:Cell Cycle; D002633:Chemotaxis / Q000235:genetics; D002874:Chromosome Mapping; D019281:Dimerization; D015964:Gene Expression Regulation, Bacterial; D016480:Helicobacter pylori / Q000235:genetics* / Q000254:growth & development; D011401:Promoter Regions, Genetic; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D015534:Trans-Activators; C112177:osmolarity response regulator proteins