Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Oct;174(19):6117–6124. doi: 10.1128/jb.174.19.6117-6124.1992

Gene structure of Enterococcus hirae (Streptococcus faecalis) F1F0-ATPase, which functions as a regulator of cytoplasmic pH.

C Shibata 1, T Ehara 1, K Tomura 1, K Igarashi 1, H Kobayashi 1
PMCID: PMC207678  PMID: 1328152

Abstract

Enterococcus hirae (formerly Streptococcus faecalis) ATCC 9790 has an F1F0-ATPase which functions as a regulator of the cytoplasmic pH but does not synthesize ATP. We isolated four clones which contained genes for c, b, delta, and alpha subunits of this enzyme but not for other subunit genes. It was revealed that two specific regions (upstream of the c-subunit gene and downstream of the gamma-subunit gene) were lost at a specific site in the clones we isolated, suggesting that these regions were unstable in Escherichia coli. The deleted regions were amplified by polymerase chain reaction, and the nucleotide sequences of these regions were determined. The results showed that eight genes for a, c, b, delta, alpha, gamma, beta, and epsilon subunits were present in this order. Northern (RNA) blot analysis showed that these eight genes were transcribed to one mRNA. The i gene was not found in the upper region of the a-subunit gene. Instead of the i gene, this operon contained a long untranslated region (240 bp) whose G + C content was only 30%. There was no typical promoter sequence such as was proposed for E. coli, suggesting that the promoter structure of this species is different from that of E. coli. Deduced amino acid sequences suggested that E. hirae H(+)-ATPase is a typical F1F0-type ATPase but that its gene structure is not identical to that of other bacterial F1F0-ATPases.

Full text

PDF
6117

Images in this article

6118Image
on p.6118
6121Image
on p.6121
6121Image
on p.6121

Selected References

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

  1. Abrams A., Jensen C. Altered expression of the H+ ATPase in Streptococcus faecalis membranes. Biochem Biophys Res Commun. 1984 Jul 18;122(1):151–157. doi: 10.1016/0006-291x(84)90452-2. [DOI] [PubMed] [Google Scholar]
  2. Abrams A., Jensen C., Morris D. H. Role of Mg2+ ions in the subunit structure and membrane binding properties of bacterial energy transducing ATPase. Biochem Biophys Res Commun. 1976 Apr 5;69(3):804–811. doi: 10.1016/0006-291x(76)90946-3. [DOI] [PubMed] [Google Scholar]
  3. Bakker E. P., Harold F. M. Energy coupling to potassium transport in Streptococcus faecalis. Interplay of ATP and the protonmotive force. J Biol Chem. 1980 Jan 25;255(2):433–440. [PubMed] [Google Scholar]
  4. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brusilow W. S., Scarpetta M. A., Hawthorne C. A., Clark W. P. Organization and sequence of the genes coding for the proton-translocating ATPase of Bacillus megaterium. J Biol Chem. 1989 Jan 25;264(3):1528–1533. [PubMed] [Google Scholar]
  6. Clarke D. J., Morris J. G. The proton-translocating adenosine triphosphatase of the obligately anaerobic bacterium Clostridium pasteurianum. 2. ATP synthetase activity. Eur J Biochem. 1979 Aug 1;98(2):613–620. doi: 10.1111/j.1432-1033.1979.tb13223.x. [DOI] [PubMed] [Google Scholar]
  7. Dillard J. P., Yother J. Analysis of Streptococcus pneumoniae sequences cloned into Escherichia coli: effect of promoter strength and transcription terminators. J Bacteriol. 1991 Aug;173(16):5105–5109. doi: 10.1128/jb.173.16.5105-5109.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  9. Franke A. E., Clewell D. B. Evidence for a chromosome-borne resistance transposon (Tn916) in Streptococcus faecalis that is capable of "conjugal" transfer in the absence of a conjugative plasmid. J Bacteriol. 1981 Jan;145(1):494–502. doi: 10.1128/jb.145.1.494-502.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fürst P., Solioz M. Formation of a beta-aspartyl phosphate intermediate by the vanadate-sensitive ATPase of Streptococcus faecalis. J Biol Chem. 1985 Jan 10;260(1):50–52. [PubMed] [Google Scholar]
  11. Harold F. M., Van Brunt J. Circulation of H+ and K+ across the plasma membrane is not obligatory for bacterial growth. Science. 1977 Jul 22;197(4301):372–373. doi: 10.1126/science.69317. [DOI] [PubMed] [Google Scholar]
  12. Heefner D. L., Harold F. M. ATP-linked sodium transport in Streptococcus faecalis. I. The sodium circulation. J Biol Chem. 1980 Dec 10;255(23):11396–11402. [PubMed] [Google Scholar]
  13. Heefner D. L., Kobayashi H., Harold F. M. ATP-linked sodium transport in Streptococcus faecalis. II. Energy coupling in everted membrane vesicles. J Biol Chem. 1980 Dec 10;255(23):11403–11407. [PubMed] [Google Scholar]
  14. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  15. Hugentobler G., Heid I., Solioz M. Purification of a putative K+-ATPase from Streptococcus faecalis. J Biol Chem. 1983 Jun 25;258(12):7611–7617. [PubMed] [Google Scholar]
  16. Kakinuma Y., Harold F. M. ATP-driven exchange of Na+ and K+ ions by Streptococcus faecalis. J Biol Chem. 1985 Feb 25;260(4):2086–2091. [PubMed] [Google Scholar]
  17. Kakinuma Y., Igarashi K. Active potassium extrusion regulated by intracellular pH in Streptococcus faecalis. J Biol Chem. 1988 Oct 5;263(28):14166–14170. [PubMed] [Google Scholar]
  18. Kanazawa H., Kiyasu T., Noumi T., Futai M. Overproduction of subunit a of the F0 component of proton-translocating ATPase inhibits growth of Escherichia coli cells. J Bacteriol. 1984 Apr;158(1):300–306. doi: 10.1128/jb.158.1.300-306.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kinoshita N., Unemoto T., Kobayashi H. Sodium-stimulated ATPase in Streptococcus faecalis. J Bacteriol. 1984 Jun;158(3):844–848. doi: 10.1128/jb.158.3.844-848.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Klionsky D. J., Brusilow W. S., Simoni R. D. In vivo evidence for the role of the epsilon subunit as an inhibitor of the proton-translocating ATPase of Escherichia coli. J Bacteriol. 1984 Dec;160(3):1055–1060. doi: 10.1128/jb.160.3.1055-1060.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kobayashi H. A proton-translocating ATPase regulates pH of the bacterial cytoplasm. J Biol Chem. 1985 Jan 10;260(1):72–76. [PubMed] [Google Scholar]
  22. Kobayashi H., Murakami N., Unemoto T. Regulation of the cytoplasmic pH in Streptococcus faecalis. J Biol Chem. 1982 Nov 25;257(22):13246–13252. [PubMed] [Google Scholar]
  23. Kobayashi H. Second system for potassium transport in Streptococcus faecalis. J Bacteriol. 1982 May;150(2):506–511. doi: 10.1128/jb.150.2.506-511.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kobayashi H., Suzuki T., Kinoshita N., Unemoto T. Amplification of the Streptococcus faecalis proton-translocating ATPase by a decrease in cytoplasmic pH. J Bacteriol. 1984 Jun;158(3):1157–1160. doi: 10.1128/jb.158.3.1157-1160.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kobayashi H., Suzuki T., Unemoto T. Streptococcal cytoplasmic pH is regulated by changes in amount and activity of a proton-translocating ATPase. J Biol Chem. 1986 Jan 15;261(2):627–630. [PubMed] [Google Scholar]
  26. Kobayashi H., Van Brunt J., Harold F. M. ATP-linked calcium transport in cells and membrane vesicles of Streptococcus faecalis. J Biol Chem. 1978 Apr 10;253(7):2085–2092. [PubMed] [Google Scholar]
  27. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  28. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  29. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  30. Leimgruber R. M., Jensen C., Abrams A. Purification and characterization of the membrane adenosine triphosphatase complex from the wild-type and N,N'-dicyclohexylcarbodiimide-resistant strains of Streptococcus faecalis. J Bacteriol. 1981 Aug;147(2):363–372. doi: 10.1128/jb.147.2.363-372.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lipman D. J., Altschul S. F., Kececioglu J. D. A tool for multiple sequence alignment. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4412–4415. doi: 10.1073/pnas.86.12.4412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J Mol Biol. 1962 Jul;5:109–118. doi: 10.1016/s0022-2836(62)80066-7. [DOI] [PubMed] [Google Scholar]
  33. Maloney P. C., Kashket E. R., Wilson T. H. A protonmotive force drives ATP synthesis in bacteria. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3896–3900. doi: 10.1073/pnas.71.10.3896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Maloney P. C., Wilson T. H. ATP synthesis driven by a protonmotive force in Streptococcus lactis. J Membr Biol. 1975;25(3-4):285–310. doi: 10.1007/BF01868580. [DOI] [PubMed] [Google Scholar]
  35. Nakayama J., Nagasawa H., Isogai A., Clewell D. B., Suzuki A. Amino acid sequence of pheromone-inducible surface protein in Enterococcus faecalis, that is encoded on the conjugative plasmid pPD1. FEBS Lett. 1990 Jul 2;267(1):81–84. doi: 10.1016/0014-5793(90)80293-r. [DOI] [PubMed] [Google Scholar]
  36. Platt T. Transcription termination and the regulation of gene expression. Annu Rev Biochem. 1986;55:339–372. doi: 10.1146/annurev.bi.55.070186.002011. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Shaw J. H., Clewell D. B. Complete nucleotide sequence of macrolide-lincosamide-streptogramin B-resistance transposon Tn917 in Streptococcus faecalis. J Bacteriol. 1985 Nov;164(2):782–796. doi: 10.1128/jb.164.2.782-796.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Suzuki T., Kobayashi H. Regulation of the cytoplasmic pH by a proton-translocating ATPase in Streptococcus faecalis (faecium). A computer simulation. Eur J Biochem. 1989 Mar 15;180(2):467–471. doi: 10.1111/j.1432-1033.1989.tb14669.x. [DOI] [PubMed] [Google Scholar]
  40. Suzuki T., Unemoto T., Kobayashi H. Novel streptococcal mutants defective in the regulation of H+-ATPase biosynthesis and in F0 complex. J Biol Chem. 1988 Aug 25;263(24):11840–11843. [PubMed] [Google Scholar]
  41. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  42. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  43. van der Drift C., Janssen D. B., van Wezenbeek P. M. Hydrolysis and synthesis of ATP by membrane-bound ATPase from a motile Streptococcus. Arch Microbiol. 1978 Oct 4;119(1):31–36. doi: 10.1007/BF00407924. [DOI] [PubMed] [Google Scholar]

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

RESOURCES