Molecular analysis of the two-component genes, ompR and envZ, in the symbiotic bacterium Xenorhabdus nematophilus.
In Escherichia coli the histidine kinase sensor protein, EnvZ, undergoes autophosphorylation and subsequently phosphorylates the regulatory protein, OmpR. Modulation of the levels of OmpR-phosphate controls the differential expression of ompF and ompC. While the phosphotransfer reaction between EnvZ and OmpR has been extensively studied, the domains involved in the sensing function of EnvZ are not well understood. We have used a comparative approach to study the sensing function of EnvZ. During our search of numerous bacteria we found that the symbiotic/pathogenic bacterium Xenorhabdus nematophilus contained the operon encoding both ompR and envZ. Nucleotide sequence analysis revealed that EnvZ of X. nematophilus (EnvZX.n.) is composed of 342 amino acid residues, which is 108 residues shorter than EnvZ of E. coli (EnvZE.c.). Amino acid sequence comparison showed that the cytoplasmic domains of the EnvZ molecules shared 57% sequence identity. In contrast, the large hydrophilic periplasmic domain of EnvZE.c. was absent in EnvZX.n., and was replaced by a shorter hydrophobic region. Although the periplasmic domains had diverged extensively, envZX.n. was able to complement a delta envZ strain of E. coli. OmpF and OmpC were differentially produced in response to changes in medium osmolarity in this strain. Further genetic analysis established that heterologous phosphorylation between EnvZX.n. and OmpR of E. coli (OmpRE.c.) accounted for the complementation of the delta envZ strain. In addition we show that the OmpR molecules of X. nematophilus and E. coli share 78% amino acid sequence identity. These results indicate that the EnvZ protein of X. nematophilus was able to sense these changes in the osmolarity of the growth environment and properly regulate the levels of OmpR-phosphate in E. coli.
Tabatabai
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Department of Biological Sciences, University of Wisconsin, Milwaukee 53201, USA.
Forst
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D000595:Amino Acid Sequence; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D004755:Enterobacteriaceae / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D005786:Gene Expression Regulation; D015723:Gene Library; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D000071677:Histidine Kinase; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D009348:Nematoda / Q000382:microbiology; D010766:Phosphorylation; D011494:Protein Kinases; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction / Q000235:genetics*; D013559:Symbiosis
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D011494:Protein Kinases; D000071677:Histidine Kinase; C057843:envZ protein, E coli