Activation of Helicobacter pylori ureA promoter by a hybrid Escherichia coli-H. pylori rpoD gene in E. coli.
We constructed and analyzed hybrid Escherichia coli-Helicobacter pylori rpoD genes in an E. coli rpoD mutant. It turned out that a hybrid consisting of E. coli rpoD with subdomain 4.2 of H. pylori rpoD (for -35 recognition) was functional. On the other hand, hybrids consisting of E. coli rpoD with domain 2 and the adjacent sequence of H. pylori rpoD (for core enzyme binding and -10 recognition) were non-functional. Intriguingly, a hybrid rpoD containing H. pylori subdomain 4.2 conferred higher activity for the H. pylori PureA as determined by xylE expression of PureA-xylE fusions, although the activity of the hybrid rpoD for the tac promoter was comparable to that of E. coli rpoD. The tsp of ureA in E. coli with the hybrid rpoD and E. coli rpoD were 15 and 17bp upstream from that in H. pylori, respectively. The comparison of PureA sequences in both E. coli and H. pylori indicated the existence of a -10 consensus sequence but little conservation of -35 sequences. Instead, the PureA in both H. pylori and E. coli contained an identical heptamer, GTTAATA, in the extended -35 region.
Shirai
M
M
Department of Microbiology, Yamaguchi University School of Medicine, Ube, Japan.
Fujinaga
R
R
Akada
J K
JK
Nakazawa
T
T
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005816:Genetic Complementation Test; D016480:Helicobacter pylori / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D012808:Sigma Factor / Q000235:genetics* / Q000502:physiology; D014158:Transcription, Genetic; D014510:Urease / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D012808:Sigma Factor; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases; D014510:Urease