Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 May;170(5):2283–2286. doi: 10.1128/jb.170.5.2283-2286.1988

Arg-220 of the PstA protein is required for phosphate transport through the phosphate-specific transport system in Escherichia coli but not for alkaline phosphatase repression.

G B Cox 1, D Webb 1, J Godovac-Zimmermann 1, H Rosenberg 1
PMCID: PMC211119  PMID: 2896188

Abstract

The pstA gene encodes an integral membrane protein of the phosphate-specific transport system of Escherichia coli. The nucleotide change in the previously described pstA2 allele was found to be a G----A substitution at position 276 of the nucleotide sequence, resulting in the premature termination of translation. Three mutations in the pstA gene were produced by site-directed mutagenesis. The amino acid substitutions resulting from the three site-directed mutations were Arg-170----Gln, Glu-173----Gln, and Arg-220----Gln. These amino acid residues were selected because a previous PstA protein structure prediction placed them within the membrane. The Arg-220----Gln mutation resulted in the loss of phosphate transport through the phosphate-specific transport system, but the alkaline phosphatase activity remained repressed. Neither the Arg-170----Gln nor the Glu-173----Gln mutation affected phosphate transport. The results are discussed in relation to a proposed structure of the PstA protein.

Full text

PDF
2283

Images in this article

2284Image
on p.2284

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amemura M., Makino K., Shinagawa H., Kobayashi A., Nakata A. Nucleotide sequence of the genes involved in phosphate transport and regulation of the phosphate regulon in Escherichia coli. J Mol Biol. 1985 Jul 20;184(2):241–250. doi: 10.1016/0022-2836(85)90377-8. [DOI] [PubMed] [Google Scholar]
  2. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bracha M., Yagil E. A ne type of alkaline phosphatase-negative mutants in Escherichia coli K12. Mol Gen Genet. 1973 Mar 27;122(1):53–60. doi: 10.1007/BF00337973. [DOI] [PubMed] [Google Scholar]
  4. Cox G. B., Rosenberg H., Downie J. A., Silver S. Genetic analysis of mutants affected in the Pst inorganic phosphate transport system. J Bacteriol. 1981 Oct;148(1):1–9. doi: 10.1128/jb.148.1.1-9.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Downie J. A., Langman L., Cox G. B., Yanofsky C., Gibson F. Subunits of the adenosine triphosphatase complex translated in vitro from the Escherichia coli unc operon. J Bacteriol. 1980 Jul;143(1):8–17. doi: 10.1128/jb.143.1.8-17.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Engelman D. M., Steitz T. A. The spontaneous insertion of proteins into and across membranes: the helical hairpin hypothesis. Cell. 1981 Feb;23(2):411–422. doi: 10.1016/0092-8674(81)90136-7. [DOI] [PubMed] [Google Scholar]
  7. Gerdes R. G., Rosenberg H. The relationship between the phosphate-binding protein and a regulator gene product from Escherichia coli. Biochim Biophys Acta. 1974 May 10;351(1):77–86. doi: 10.1016/0005-2795(74)90066-x. [DOI] [PubMed] [Google Scholar]
  8. Higgins C. F., Haag P. D., Nikaido K., Ardeshir F., Garcia G., Ames G. F. Complete nucleotide sequence and identification of membrane components of the histidine transport operon of S. typhimurium. Nature. 1982 Aug 19;298(5876):723–727. doi: 10.1038/298723a0. [DOI] [PubMed] [Google Scholar]
  9. Jans D. A., Fimmel A. L., Langman L., James L. B., Downie J. A., Senior A. E., Ash G. R., Gibson F., Cox G. B. Mutations in the uncE gene affecting assembly of the c-subunit of the adenosine triphosphatase of Escherichia coli. Biochem J. 1983 Jun 1;211(3):717–726. doi: 10.1042/bj2110717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lightowlers R. N., Howitt S. M., Hatch L., Gibson F., Cox G. B. The proton pore in the Escherichia coli F0F1-ATPase: a requirement for arginine at position 210 of the a-subunit. Biochim Biophys Acta. 1987 Dec 17;894(3):399–406. doi: 10.1016/0005-2728(87)90118-6. [DOI] [PubMed] [Google Scholar]
  11. Rosenberg H., Cox G. B., Butlin J. D., Gutowski S. J. Metabolite transport in mutants of Escherichia coli K12 defective in electron transport and coupled phosphorylation. Biochem J. 1975 Feb;146(2):417–423. doi: 10.1042/bj1460417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rosenberg H. Transport of iron into bacterial cells. Methods Enzymol. 1979;56:388–394. doi: 10.1016/0076-6879(79)56036-4. [DOI] [PubMed] [Google Scholar]
  13. SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
  14. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Selker E., Brown K., Yanofsky C. Mitomycin C-induced expression of trpA of Salmonella typhimurium inserted into the plasmid ColE1. J Bacteriol. 1977 Jan;129(1):388–394. doi: 10.1128/jb.129.1.388-394.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sprague G. F., Jr, Bell R. M., Cronan J. E., Jr A mutant of Escherichia coli auxotrophic for organic phosphates: evidence for two defects in inorganic phosphate transport. Mol Gen Genet. 1975 Dec 30;143(1):71–77. doi: 10.1007/BF00269422. [DOI] [PubMed] [Google Scholar]
  17. Surin B. P., Jans D. A., Fimmel A. L., Shaw D. C., Cox G. B., Rosenberg H. Structural gene for the phosphate-repressible phosphate-binding protein of Escherichia coli has its own promoter: complete nucleotide sequence of the phoS gene. J Bacteriol. 1984 Mar;157(3):772–778. doi: 10.1128/jb.157.3.772-778.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Surin B. P., Rosenberg H., Cox G. B. Phosphate-specific transport system of Escherichia coli: nucleotide sequence and gene-polypeptide relationships. J Bacteriol. 1985 Jan;161(1):189–198. doi: 10.1128/jb.161.1.189-198.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zuckier G., Torriani A. Genetic and physiological tests of three phosphate-specific transport mutants of Escherichia coli. J Bacteriol. 1981 Mar;145(3):1249–1256. doi: 10.1128/jb.145.3.1249-1256.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES