Molecular characterization of HPr and related enzymes, and regulation of HPr phosphorylation in the ruminal bacterium Streptococcus bovis.
Molecular properties of HPr, enzyme I, and HPr kinase in Streptococcus bovis, and the regulation of HPr phosphorylation were examined. The genes encoding HPr (ptsH) and enzyme I (ptsI) were found to be cotranscribed. Two transcriptional start sites were detected in a region upstream of the HPr kinase gene (hprK). HPr kinase had both HPr-phosphorylating and HPr-dephosphorylating activities. The importance of phosphorylation of Ser-46 in HPr was shown by using a mutant HPr in which Ser-46 was replaced by Ala. When S. bovis was grown in glucose-limited medium, the amount of seryl-phosphorylated HPr (HPr-[Ser-P]) decreased drastically as the growth rate decreased. In contrast, the amount of histidyl-phosphorylated HPr (HPr-[His-P]) increased gradually as the growth rate decreased. The amount of HPr kinase did not greatly change with the growth phase, whereas the intracellular P(i) concentration increased as the growth rate decreased. HPr-[Ser-P] decreased as the intracellular P(i) increased as a consequence of inhibition of HPr kinase activity by P(i) and simultaneous enhancement of HPr-[Ser-P] phosphatase activity by P(i). Thus, it is conceivable that the ratio of HPr-[Ser-P] to HPr-[His-P] is regulated by the bifunctional activity of HPr kinase in response to intracellular P(i) concentration.
Asanuma
Narito
N
Department of Life Science, College of Agriculture, Meiji University, Higashimita, Tama-ku, 214-8571 Kawasaki, Japan.
Hino
Tsuneo
T
D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010766:Phosphorylation; D017852:Phosphotransferases (Nitrogenous Group Acceptor) / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D012417:Rumen / Q000382:microbiology; D016985:Streptococcus bovis / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C030182:phosphocarrier protein HPr; C110847:HPr kinase; D017346:Protein Serine-Threonine Kinases; D017852:Phosphotransferases (Nitrogenous Group Acceptor); C022360:phosphoenolpyruvate-protein phosphotransferase