Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1983 Sep;155(3):1265–1270. doi: 10.1128/jb.155.3.1265-1270.1983

Cloning and expression of uncI, the first gene of the unc operon of Escherichia coli.

W S Brusilow, A C Porter, R D Simoni
PMCID: PMC217824  PMID: 6224772

Abstract

The unc operon of Escherichia coli consists of eight genes coding for the eight subunits of the proton-translocating ATPase. In vitro transcription-translation of DNA cloned from the beginning of the operon onto plasmids reveals that the reading frame uncI, which precedes the other genes of the operon, codes for a protein with a molecular weight of 14,500, called i. In minicells, the i protein is synthesized in amounts comparable to the amounts of the ATPase subunits, suggesting that it may be part of the ATPase complex. The presence of the unc promoter and uncI on a plasmid containing the other eight genes of the unc operon has little effect on the differential expression of the unc genes or the partitioning of the newly synthesized subunits into soluble or sedimentable fractions in the in vitro system. The i protein partitions into the sedimentable fraction.

Full text

PDF
1265

Images in this article

1266Image
on p.1266
1267Image
on p.1267
1268Image
on p.1268

Selected References

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

  1. Brusilow W. S., Gunsalus R. P., Hardeman E. C., Decker K. P., Simoni R. D. In vitro synthesis of the F0 and F1 components of the proton translocating ATPase of Escherichia coli. J Biol Chem. 1981 Apr 10;256(7):3141–3144. [PubMed] [Google Scholar]
  2. Brusilow W. S., Klionsky D. J., Simoni R. D. Differential polypeptide synthesis of the proton-translocating ATPase of Escherichia coli. J Bacteriol. 1982 Sep;151(3):1363–1371. doi: 10.1128/jb.151.3.1363-1371.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Decker K. P., Brusilow W. S., Gunsalus R. P., Simoni R. D. In vitro membrane association of the F0 polypeptides of the Escherichia coli proton translocating ATPase. J Bacteriol. 1982 Nov;152(2):815–821. doi: 10.1128/jb.152.2.815-821.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dougan G., Sherratt D. The transposon Tn1 as a probe for studying ColE1 structure and function. Mol Gen Genet. 1977 Mar 7;151(2):151–160. doi: 10.1007/BF00338689. [DOI] [PubMed] [Google Scholar]
  5. Downie J. A., Cox G. B., Langman L., Ash G., Becker M., Gibson F. Three genes coding for subunits of the membrane sector (F0) of the Escherichia coli adenosine triphosphatase complex. J Bacteriol. 1981 Jan;145(1):200–210. doi: 10.1128/jb.145.1.200-210.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Downie J. A., Gibson F., Cox G. B. Membrane adenosine triphosphatases of prokaryotic cells. Annu Rev Biochem. 1979;48:103–131. doi: 10.1146/annurev.bi.48.070179.000535. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Gay N. J., Walker J. E. The atp operon: nucleotide sequence of the promoter and the genes for the membrane proteins, and the delta subunit of Escherichia coli ATP-synthase. Nucleic Acids Res. 1981 Aug 25;9(16):3919–3926. doi: 10.1093/nar/9.16.3919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gay N. J., Walker J. E. The atp operon: nucleotide sequence of the region encoding the alpha-subunit of Escherichia coli ATP-synthase. Nucleic Acids Res. 1981 May 11;9(9):2187–2194. doi: 10.1093/nar/9.9.2187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gunsalus R. P., Brusilow W. S., Simoni R. D. Gene order and gene-polypeptide relationships of the proton-translocating ATPase operon (unc) of Escherichia coli. Proc Natl Acad Sci U S A. 1982 Jan;79(2):320–324. doi: 10.1073/pnas.79.2.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gunsalus R. P., Zurawski G., Yanofsky C. Structural and functional analysis of cloned deoxyribonucleic acid containing the trpR-thr region of the Escherichia coli chromosome. J Bacteriol. 1979 Oct;140(1):106–113. doi: 10.1128/jb.140.1.106-113.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hansen F. G., Nielsen J., Riise E., von Meyenburg K. The genes for the eight subunits of the membrane bound ATP synthase of Escherichia coli. Mol Gen Genet. 1981;183(3):463–472. doi: 10.1007/BF00268766. [DOI] [PubMed] [Google Scholar]
  13. Harding N. E., Cleary J. M., Smith D. W., Michon J. J., Brusilow W. S., Zyskind J. W. Chromosomal replication origins (oriC) of Enterobacter aerogenes and Klebsiella pneumoniae are functional in Escherichia coli. J Bacteriol. 1982 Dec;152(3):983–993. doi: 10.1128/jb.152.3.983-993.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kanazawa H., Mabuchi K., Futai M. Nucleotide sequence of the promoter region of the gene cluster for proton-translocating ATPase from Escherichia coli and identification of the active promotor. Biochem Biophys Res Commun. 1982 Jul 30;107(2):568–575. doi: 10.1016/0006-291x(82)91529-7. [DOI] [PubMed] [Google Scholar]
  15. Kanazawa H., Mabuchi K., Kayano T., Noumi T., Sekiya T., Futai M. Nucleotide sequence of the genes for F0 components of the proton-translocating ATPase from Escherichia coli: prediction of the primary structure of F0 subunits. Biochem Biophys Res Commun. 1981 Nov 30;103(2):613–620. doi: 10.1016/0006-291x(81)90495-2. [DOI] [PubMed] [Google Scholar]
  16. Klionsky D. J., Brusilow W. S., Simoni R. D. Assembly of a functional F0 of the proton-translocating ATPase of Escherichia coli. J Biol Chem. 1983 Aug 25;258(16):10136–10143. [PubMed] [Google Scholar]
  17. Mabuchi K., Kanazawa H., Kayano T., Futai M. Nucleotide sequence of the gene coding for the delta subunit of proton translocating ATPase of Escherichia coli. Biochem Biophys Res Commun. 1981 Sep 16;102(1):172–179. doi: 10.1016/0006-291x(81)91504-7. [DOI] [PubMed] [Google Scholar]
  18. Ogura T., Miki T., Hiraga S. Copy-number mutants of the plasmid carrying the replication origin of the Escherichia coli chromosome: evidence for a control region of replication. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3993–3997. doi: 10.1073/pnas.77.7.3993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Porter A. C., Brusilow W. S., Simoni R. D. Promoter for the unc operon of Escherichia coli. J Bacteriol. 1983 Sep;155(3):1271–1278. doi: 10.1128/jb.155.3.1271-1278.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Roozen K. J., Fenwick R. G., Jr, Curtiss R., 3rd Synthesis of ribonucleic acid and protein in plasmid-containing minicells of Escherichia coli K-12. J Bacteriol. 1971 Jul;107(1):21–33. doi: 10.1128/jb.107.1.21-33.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Saraste M., Gay N. J., Eberle A., Runswick M. J., Walker J. E. The atp operon: nucleotide sequence of the genes for the gamma, beta, and epsilon subunits of Escherichia coli ATP synthase. Nucleic Acids Res. 1981 Oct 24;9(20):5287–5296. doi: 10.1093/nar/9.20.5287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yamaguchi K., Yamaguchi M. Copy number mutations (Cop-) of the plasmid containing the replication origin (oriC) of the Escherichia coli chromosome: lethal effect of the cop region cloned onto a high-copy-number vector on host cells. J Bacteriol. 1983 Jan;153(1):550–554. doi: 10.1128/jb.153.1.550-554.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. von Meyenburg K., Jørgensen B. B., Nielsen J., Hansen F. G. Promoters of the atp operon coding for the membrane-bound ATP synthase of Escherichia coli mapped by Tn10 insertion mutations. Mol Gen Genet. 1982;188(2):240–248. doi: 10.1007/BF00332682. [DOI] [PubMed] [Google Scholar]

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

RESOURCES