Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1981 Jan;145(1):200–210. doi: 10.1128/jb.145.1.200-210.1981

Three genes coding for subunits of the membrane sector (F0) of the Escherichia coli adenosine triphosphatase complex.

J A Downie, G B Cox, L Langman, G Ash, M Becker, F Gibson
PMCID: PMC217262  PMID: 6450744

Abstract

Two mutant unc alleles, unc-469 and unc-476, have been characterized as affecting a previously undescribed gene, designated uncF. The uncF gene is part of the unc operon (with the gene order being uncBFEAGDC), although some uncertainty remains as to the relative order of the uncF and uncE genes. Mutant strains carrying the uncF469 or uncF476 allele lack the 18,000-molecular-weight component of the F0 sector of the adenosine triphosphatase in the cell membrane but retain the dicyclohexylcarbodiimide-binding protein (molecular weight, 8,400). Conversely, strains carrying mutations in the uncE gene lack the dicyclohexylcarbodiimide-binding protein but retain the 18,000-molecular-weight protein in the cell membrane. Strains carrying mutations in the uncB gene have both the 18,000-molecular-weight protein and the dicyclohexylcarbodiimide-binding protein present in the cell membranes. The three proteins of the F0 portion of the adenosine triphosphatase, viz., 24,000, 18,000, and 8,400 molecular weights, became membrane associated after in vitro transcription-translation with plasmid pAN51 as template. Plasmids carrying deletions which affected the UncBFE region were isolated from plasmid pAN51 and characterized genetically. A comparison of the genes that were absent from the various deletion plasmids with the membrane-associated products formed after in vitro transcription-translation indicated that the uncB gene coded for the 24,000-molecular-weight protein and that the gene order was probably uncBFE. A correlation between length of deoxyribonucleic acid, genes present, and their products is presented in relation to plasmid pAN51.

Full text

PDF

Images in this article

Selected References

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

  1. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barnes W. M. Plasmid detection and sizing in single colony lysates. Science. 1977 Jan 28;195(4276):393–394. doi: 10.1126/science.318764. [DOI] [PubMed] [Google Scholar]
  3. Butlin J. D., Cox G. B., Gibson F. Oxidative phosphorylation in Escherichia coli K-12: the genetic and biochemical characterisations of a strain carrying a mutation in the uncB gene. Biochim Biophys Acta. 1973 Feb 22;292(2):366–375. doi: 10.1016/0005-2728(73)90043-1. [DOI] [PubMed] [Google Scholar]
  4. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cox G. B., Downie J. A., Fayle D. R., Gibson F., Radik J. Inhibition, by a protease inhibitor, of the solubilization of the F1-portion of the Mg2+-stimulated adenosine triphosphatase of Escherichia coli. J Bacteriol. 1978 Jan;133(1):287–292. doi: 10.1128/jb.133.1.287-292.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cox G. B., Downie J. A., Gibson F., Radik J. Genetic complementation between two mutant unc alleles (unc A401 and unc D409) affecting the Fl portion of the magnesium ion-stimulated adenosine triphosphatase of Escherichia coli K12. Biochem J. 1978 Mar 15;170(3):593–598. doi: 10.1042/bj1700593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cox G. B., Downie J. A. Isolation and characterization of mutants of Escherichia coli K-12 affected in oxidative phosphorylation of quinone biosynthesis. Methods Enzymol. 1979;56:106–117. doi: 10.1016/0076-6879(79)56013-3. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Downie J. A., Senior A. E., Gibson F., Cox G. B. A fifth gene (uncE) in the operon concerned with oxidative phosphorylation in Escherichia coli. J Bacteriol. 1979 Feb;137(2):711–718. doi: 10.1128/jb.137.2.711-718.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dunn S. D. Identification of the altered subunit in the inactive F1ATPase of an Escherichia coli uncA mutant. Biochem Biophys Res Commun. 1978 May 30;82(2):596–602. doi: 10.1016/0006-291x(78)90916-6. [DOI] [PubMed] [Google Scholar]
  11. Fayle D. R., Downie J. A., Cox G. B., Gibson F., Radik J. Characterization of the mutant-unc D-gene product in a strain of Escherichia coli K12. An altered beta-subunit of the magnesium ion-stimulated adenosine triphosphatase. Biochem J. 1978 Jun 15;172(3):523–531. doi: 10.1042/bj1720523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fillingame R. H. Identification of the dicyclohexylcarbodiimide-reactive protein component of the adenosine 5'-triphosphate energy-transducing system of Escherichia coli. J Bacteriol. 1975 Nov;124(2):870–883. doi: 10.1128/jb.124.2.870-883.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fillingame R. H. Purification of the carbodiimide-reactive protein component of the ATP energy-transducing system of Escherichia coli. J Biol Chem. 1976 Nov 10;251(21):6630–6637. [PubMed] [Google Scholar]
  14. Foster D. L., Fillingame R. H. Energy-transducing H+-ATPase of Escherichia coli. Purification, reconstitution, and subunit composition. J Biol Chem. 1979 Sep 10;254(17):8230–8236. [PubMed] [Google Scholar]
  15. Friedl P., Friedl C., Schairer H. U. The ATP synthetase of Escherichia coli K12: purification of the enzyme and reconstitution of energy-transducing activities. Eur J Biochem. 1979 Oct;100(1):175–180. doi: 10.1111/j.1432-1033.1979.tb02046.x. [DOI] [PubMed] [Google Scholar]
  16. Gibson F., Cox G. B., Downie J. A., Radik J. A mutation affecting a second component of the F0 portion of the magnesium ion-stimulated adenosine triphosphatase of Escherichia coli K12. The uncC424 allele. Biochem J. 1977 Apr 15;164(1):193–198. doi: 10.1042/bj1640193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gibson F., Cox G. B., Downie J. A., Radik J. Partial diploids of Escherichia coli carrying normal and mutant alleles affecting oxidative phosphorylation. Biochem J. 1977 Mar 15;162(3):665–670. doi: 10.1042/bj1620665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gibson F., Downie J. A., Cox G. B., Radik J. Mu-induced polarity in the unc operon of Escherichia coli. J Bacteriol. 1978 Jun;134(3):728–736. doi: 10.1128/jb.134.3.728-736.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Haddock B. A., Downie J. A. The reconstitution of functional respiratory chains in membranes from electron-transport-deficient mutants of Escherichia coli as demonstrated by quenching of atebrin fluorescence. Biochem J. 1974 Sep;142(3):703–706. doi: 10.1042/bj1420703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Haddock B. A., Jones C. W. Bacterial respiration. Bacteriol Rev. 1977 Mar;41(1):47–99. doi: 10.1128/br.41.1.47-99.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hare J. F. Purification and characterization of a dicyclohexylcarbodiimide-sensitive adenosine triphosphatase complex from membranes of Escherichia coli. Biochem Biophys Res Commun. 1975 Oct 27;66(4):1329–1337. doi: 10.1016/0006-291x(75)90505-7. [DOI] [PubMed] [Google Scholar]
  22. Kanazawa H., Saito S., Futai M. Coupling factor ATPase from Escherichia coli. An uncA mutant (uncA401) with defective alpha subunit. J Biochem. 1978 Dec;84(6):1513–1517. doi: 10.1093/oxfordjournals.jbchem.a132276. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Lederberg E. M., Cohen S. N. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. doi: 10.1128/jb.119.3.1072-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Messer W., Meijer M., Bergmans H. E., Hansen F. G., von Meyenburg K., Beck E., Schaller H. Origin of replication, oriC, of the Escherichia coli K12 chromosome: nucleotide sequence. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):139–145. doi: 10.1101/sqb.1979.043.01.020. [DOI] [PubMed] [Google Scholar]
  26. Novick R. P., Clowes R. C., Cohen S. N., Curtiss R., 3rd, Datta N., Falkow S. Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev. 1976 Mar;40(1):168–189. doi: 10.1128/br.40.1.168-189.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  28. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  29. Rosen B. P., Hasan S. M. Purification of an N,N'-dicyclohexylcarbodiimide-sensitive ATPase from Escherichia coli. FEBS Lett. 1979 Aug 15;104(2):339–342. doi: 10.1016/0014-5793(79)80847-9. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Senior A. E., Downie J. A., Cox G. B., Gibson F., Langman L., Fayle D. R. The uncA gene codes for the alpha-subunit of the adenosine triphosphatase of Escherichia coli. Electrophoretic analysis of uncA mutant strains. Biochem J. 1979 Apr 15;180(1):103–109. doi: 10.1042/bj1800103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Senior A. E., Fayle D. R., Downie J. A., Gibson F., Cox G. B. Properties of membranes from mutant strains of Escherichia coli in which the beta-subunit of the adenosine triphosphatase is abnormal. Biochem J. 1979 Apr 15;180(1):111–118. doi: 10.1042/bj1800111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Takahashi H., Shimizu M., Saito H., Ikeda Y. Studies of viable T4 bacteriophage containing cytosine-substituted DNA (T4dC phage). II. Cleavage of T4dC DNA by endonuclease SalI and bam HI. Mol Gen Genet. 1979 Jan 5;168(1):49–53. doi: 10.1007/BF00267932. [DOI] [PubMed] [Google Scholar]
  34. Zalkin H., Yanofsky C., Squires C. L. Regulated in vitro synthesis of Escherichia coli tryptophan operon messenger ribonucleic acid and enzymes. J Biol Chem. 1974 Jan 25;249(2):465–475. [PubMed] [Google Scholar]

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

RESOURCES