Abstract
We investigated the regulation of the synthesis of the eight polypeptides of the Escherichia coli proton-translocating ATPase. A plasmid carrying the eight genes of the unc operon was used to direct in vivo and in vitro protein synthesis of the eight polypeptides. Analysis of these data indicates that the ATPase polypeptides are synthesized in unequal amounts both in vitro and in vivo. We identified several regions within the unc operon at which expression of a gene is either increased or decreased from that of the preceding gene. Since genetic information indicates a single polycistronic mRNA for all eight genes of this operon, the observed differential synthesis of the polypeptides is most likely the result of translational regulation. The effect of varying the temperature suggests that the secondary structure in the mRNA may affect the rate of translation initiation in the region between uncE and uncF.
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.
- Borer P. N., Dengler B., Tinoco I., Jr, Uhlenbeck O. C. Stability of ribonucleic acid double-stranded helices. J Mol Biol. 1974 Jul 15;86(4):843–853. doi: 10.1016/0022-2836(74)90357-x. [DOI] [PubMed] [Google Scholar]
- Bragg P. D., Hou C. Purification of a factor for both aerobic-driven and ATP-driven energy-dependent transhydrogenases of Escherichia coli. FEBS Lett. 1972 Dec 15;28(3):309–312. doi: 10.1016/0014-5793(72)80738-5. [DOI] [PubMed] [Google Scholar]
- 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]
- DeFranco A. L., Koshland D. E., Jr Molecular cloning of chemotaxis genes and overproduction of gene products in the bacterial sensing system. J Bacteriol. 1981 Aug;147(2):390–400. doi: 10.1128/jb.147.2.390-400.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- Dunn J. J., Buzash-Pollert E., Studier F. W. Mutations of bacteriophage T7 that affect initiation of synthesis of the gene 0.3 protein. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2741–2745. doi: 10.1073/pnas.75.6.2741. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Foster D. L., Fillingame R. H. Stoichiometry of subunits in the H+-ATPase complex of Escherichia coli. J Biol Chem. 1982 Feb 25;257(4):2009–2015. [PubMed] [Google Scholar]
- 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]
- Friedl P., Schairer H. U. The isolated F0 of Escherichia coli aTP-synthase is reconstitutively active in H+-conduction and ATP-dependent energy-transduction. FEBS Lett. 1981 Jun 15;128(2):261–264. doi: 10.1016/0014-5793(81)80094-4. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Gold L., Pribnow D., Schneider T., Shinedling S., Singer B. S., Stormo G. Translational initiation in prokaryotes. Annu Rev Microbiol. 1981;35:365–403. doi: 10.1146/annurev.mi.35.100181.002053. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Kanazawa H., Kayano T., Mabuchi K., Futai M. Nucleotide sequence of the genes coding for alpha, beta and gamma subunits of the proton-translocating ATPase of Escherichia coli. Biochem Biophys Res Commun. 1981 Nov 30;103(2):604–612. doi: 10.1016/0006-291x(81)90494-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Kanazawa H., Miki T., Tamura F., Yura T., Futai M. Specialized transducing phage lambda carrying the genes for coupling factor of oxidative phosphorylation of Escherichia coli: increased synthesis of coupling factor on induction of prophage lambda asn. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1126–1130. doi: 10.1073/pnas.76.3.1126. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Nielsen J., Hansen F. G., Hoppe J., Friedl P., von Meyenburg K. The nucleotide sequence of the atp genes coding for the F0 subunits a, b, c and the F1 subunit delta of the membrane bound ATP synthase of Escherichia coli. Mol Gen Genet. 1981;184(1):33–39. doi: 10.1007/BF00271191. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simoni R. D., Shandell A. Energy transduction in Escherichia coli. Genetic alteration of a membrane polypeptide of the (Ca2+,Mg2+)-ATPase. J Biol Chem. 1975 Dec 25;250(24):9421–9427. [PubMed] [Google Scholar]
- Steege D. A. 5'-Terminal nucleotide sequence of Escherichia coli lactose repressor mRNA: features of translational initiation and reinitiation sites. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4163–4167. doi: 10.1073/pnas.74.10.4163. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steitz J. A., Jakes K. How ribosomes select initiator regions in mRNA: base pair formation between the 3' terminus of 16S rRNA and the mRNA during initiation of protein synthesis in Escherichia coli. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4734–4738. doi: 10.1073/pnas.72.12.4734. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
- Yates J. L., Arfsten A. E., Nomura M. In vitro expression of Escherichia coli ribosomal protein genes: autogenous inhibition of translation. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1837–1841. doi: 10.1073/pnas.77.4.1837. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]

