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PMID: 7961438 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Bacillus subtilis F0F1 ATPase: DNA sequence of the atp operon and characterization of atp mutants.

Journal of bacteriology ·Vol. 176 ·No. 22 ·1994-11-00 ·Pages 6802-11

Santana M, Ionescu MS, Vertes A, Longin R, Kunst F, Danchin A, Glaser P

Abstract

We cloned and sequenced an operon of nine genes coding for the subunits of the Bacillus subtilis F0F1 ATP synthase. The arrangement of these genes in the operon is identical to that of the atp operon from Escherichia coli and from three other Bacillus species. The deduced amino acid sequences of the nine subunits are very similar to their counterparts from other organisms. We constructed two B. subtilis strains from which different parts of the atp operon were deleted. These B. subtilis atp mutants were unable to grow with succinate as the sole carbon and energy source. ATP was synthesized in these strains only by substrate-level phosphorylation. The two mutants had a decreased growth yield (43 and 56% of the wild-type level) and a decreased growth rate (61 and 66% of the wild-type level), correlating with a twofold decrease of the intracellular ATP/ADP ratio. In the absence of oxidative phosphorylation, B. subtilis increased ATP synthesis through substrate-level phosphorylation, as shown by the twofold increase of by-product formation (mainly acetate). The increased turnover of glycolysis in the mutant strain presumably led to increased synthesis of NADH, which would account for the observed stimulation of the respiration rate associated with an increase in the expression of genes coding for respiratory enzymes. It therefore appears that B. subtilis and E. coli respond in similar ways to the absence of oxidative phosphorylation.

MeSH Terms
Adenosine Diphosphate/analysis Adenosine Triphosphate/analysis Amino Acid Sequence Bacillus subtilis/enzymology,genetics,growth & development Base Sequence Carbon/metabolism Cloning, Molecular Energy Metabolism/genetics Genes, Bacterial/genetics Glycolysis Molecular Sequence Data Mutagenesis Operon/genetics Oxidative Phosphorylation Oxidoreductases/genetics Proton-Translocating ATPases/biosynthesis,genetics Recombinant Fusion Proteins Restriction Mapping Sequence Analysis, DNA Sequence Deletion Sequence Homology, Amino Acid Transcription, Genetic
Chemicals
Recombinant Fusion Proteins Adenosine Diphosphate Carbon Adenosine Triphosphate Oxidoreductases Proton-Translocating ATPases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Santana M
Unité de Régulation de l'Expression Génétique, Institut Pasteur, Paris, France.
Ionescu M S
Vertes A
Longin R
Kunst F
Danchin A
Glaser P
References (56)
56 references, click to expand
  1. Catabolic repression of bacterial sporulation.
    Proc Natl Acad Sci U S A. 1965 Sep;54(3):704-11 PMID: 4956288
  2. Bacillus subtilis genome project: cloning and sequencing of the 97 kb region from 325 degrees to 333 degrees.
    Mol Microbiol. 1993 Oct;10(2):371-84 PMID: 7934828
  3. Restoration of active transport in an Mg2+-adenosine triphosphatase-deficient mutant of Escherichia coli.
    J Bacteriol. 1973 Dec;116(3):1124-9 PMID: 4270946
  4. Use of neomycin in the isolation of mutants blocked in energy conservation in Escherichia coli.
    J Bacteriol. 1972 Jul;111(1):287-9 PMID: 4273171
  5. Revision of the linkage map of Bacillus subtilis 168: indications for circularity of the chromosome.
    J Bacteriol. 1975 Mar;121(3):823-34 PMID: 803963
  6. Subunit composition, function, and spatial arrangement in the Ca2+-and Mg2+-activated adenosine triphosphatases of Escherichia coli and Salmonella typhimurium.
    Arch Biochem Biophys. 1975 Mar;167(1):311-21 PMID: 124154
  7. Identification of the dicyclohexylcarbodiimide-reactive protein component of the adenosine 5'-triphosphate energy-transducing system of Escherichia coli.
    J Bacteriol. 1975 Nov;124(2):870-83 PMID: 126994
  8. Patterns of protein synthesis in E. coli: a catalog of the amount of 140 individual proteins at different growth rates.
    Cell. 1978 May;14(1):179-90 PMID: 352533
  9. Reconstitution of the energy transformer, gate and channel subunit reassembly, crystalline ATPase and ATP synthesis.
    Biochim Biophys Acta. 1978 Sep 21;505(1):45-93 PMID: 30482
  10. Energy-transducing H+-ATPase of Escherichia coli. Purification, reconstitution, and subunit composition.
    J Biol Chem. 1979 Sep 10;254(17):8230-6 PMID: 38249
  11. Membrane adenosine triphosphatases of prokaryotic cells.
    Annu Rev Biochem. 1979;48:103-31 PMID: 157712
  12. Identification of common molecular subsequences.
    J Mol Biol. 1981 Mar 25;147(1):195-7 PMID: 7265238
  13. 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):3919-26 PMID: 6272190
  14. Stoichiometry of subunits in the H+-ATPase complex of Escherichia coli.
    J Biol Chem. 1982 Feb 25;257(4):2009-15 PMID: 6460031
  15. 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):613-20 PMID: 6277311
  16. The genes for the eight subunits of the membrane bound ATP synthase of Escherichia coli.
    Mol Gen Genet. 1981;183(3):463-72 PMID: 6460910
  17. Nucleotide sequence and functional map of pC194, a plasmid that specifies inducible chloramphenicol resistance.
    J Bacteriol. 1982 May;150(2):815-25 PMID: 6950931
  18. Solid-phase sequence analysis of polypeptides eluted from polyacrylamide gels. An aid to interpretation of DNA sequences exemplified by the Escherichia coli unc operon and bacteriophage lambda.
    Eur J Biochem. 1982 Apr 1;123(2):253-60 PMID: 6210528
  19. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments.
    Gene. 1982 Oct;19(3):269-76 PMID: 6295880
  20. Insertional mutagenesis in Bacillus subtilis: mechanism and use in gene cloning.
    Gene. 1982 Oct;19(3):277-84 PMID: 6295881
  21. 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):240-8 PMID: 6185823
  22. Studies on transformation of Escherichia coli with plasmids.
    J Mol Biol. 1983 Jun 5;166(4):557-80 PMID: 6345791
  23. Nucleotide sequence of the Streptococcus faecalis plasmid gene encoding the 3'5"-aminoglycoside phosphotransferase type III.
    Gene. 1983 Sep;23(3):331-41 PMID: 6313476
  24. Physiological and morphological effects of overproduction of membrane-bound ATP synthase in Escherichia coli K-12.
    EMBO J. 1984 Aug;3(8):1791-7 PMID: 6090127
  25. Rhodopseudomonas blastica atp operon. Nucleotide sequence and transcription.
    J Mol Biol. 1984 Oct 25;179(2):185-214 PMID: 6209404
  26. DNA sequence around the Escherichia coli unc operon. Completion of the sequence of a 17 kilobase segment containing asnA, oriC, unc, glmS and phoS.
    Biochem J. 1984 Dec 15;224(3):799-815 PMID: 6395859
  27. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  28. Nucleotide sequence of the Rhodospirillum rubrum atp operon.
    Biochem J. 1985 Jun 1;228(2):391-407 PMID: 2861810
  29. Translational initiation frequency of atp genes from Escherichia coli: identification of an intercistronic sequence that enhances translation.
    EMBO J. 1985 Feb;4(2):519-26 PMID: 2862030
  30. Genes encoding the beta and epsilon subunits of the proton-translocating ATPase from Anabaena sp. strain PCC 7120.
    J Bacteriol. 1987 Jan;169(1):80-6 PMID: 2878921
  31. The organization and sequence of the genes for ATP synthase subunits in the cyanobacterium Synechococcus 6301. Support for an endosymbiotic origin of chloroplasts.
    J Mol Biol. 1987 Apr 5;194(3):359-83 PMID: 3041005
  32. Sequence and over-expression of subunits of adenosine triphosphate synthase in thermophilic bacterium PS3.
    Biochim Biophys Acta. 1988 Mar 30;933(1):141-55 PMID: 2894854
  33. 'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers.
    Nucleic Acids Res. 1988 Mar 11;16(5):1829-36 PMID: 2832831
  34. Improved tools for biological sequence comparison.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444-8 PMID: 3162770
  35. A rapid and convenient method for the preparation and storage of competent bacterial cells.
    Nucleic Acids Res. 1988 Apr 25;16(8):3580 PMID: 3287331
  36. Organization and sequence of the genes coding for the proton-translocating ATPase of Bacillus megaterium.
    J Biol Chem. 1989 Jan 25;264(3):1528-33 PMID: 2521483
  37. The pMTL nic- cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing.
    Gene. 1988 Aug 15;68(1):139-49 PMID: 2851488
  38. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer.
    Gene. 1988 Dec 15;73(1):237-44 PMID: 3243435
  39. Bacillus subtilis CtaA and CtaB function in haem A biosynthesis.
    Mol Microbiol. 1993 Oct;10(1):193-201 PMID: 7968515
  40. Signal transduction pathway controlling synthesis of a class of degradative enzymes in Bacillus subtilis: expression of the regulatory genes and analysis of mutations in degS and degU.
    J Bacteriol. 1990 Feb;172(2):824-34 PMID: 1688843
  41. Basic local alignment search tool.
    J Mol Biol. 1990 Oct 5;215(3):403-10 PMID: 2231712
  42. The Bacillus subtilis cytochrome-c oxidase. Variations on a conserved protein theme.
    Eur J Biochem. 1991 Jan 30;195(2):517-25 PMID: 1847686
  43. DegS-DegU and ComP-ComA modulator-effector pairs control expression of the Bacillus subtilis pleiotropic regulatory gene degQ.
    J Bacteriol. 1991 Apr;173(7):2366-77 PMID: 1901055
  44. A sequence assembly and editing program for efficient management of large projects.
    Nucleic Acids Res. 1991 Jul 25;19(14):3907-11 PMID: 1861983
  45. Complete physical map of the Bacillus subtilis 168 chromosome constructed by a gene-directed mutagenesis method.
    J Mol Biol. 1991 Aug 5;220(3):631-48 PMID: 1908013
  46. Organization and nucleotide sequence of the atp genes encoding the ATP synthase from alkaliphilic Bacillus firmus OF4.
    Mol Gen Genet. 1991 Oct;229(2):292-300 PMID: 1833620
  47. The project of sequencing the entire Bacillus subtilis genome.
    Res Microbiol. 1991 Sep-Oct;142(7-8):905-12 PMID: 1784829
  48. Differential gene expression from the Escherichia coli atp operon mediated by segmental differences in mRNA stability.
    Mol Microbiol. 1991 Oct;5(10):2447-58 PMID: 1838784
  49. Multiple IS insertion sequences near the replication terminus in Escherichia coli K-12.
    Biochimie. 1991 Nov;73(11):1361-74 PMID: 1665988
  50. Molecular cloning, sequencing, and physiological characterization of the qox operon from Bacillus subtilis encoding the aa3-600 quinol oxidase.
    J Biol Chem. 1992 May 25;267(15):10225-31 PMID: 1316894
  51. Nucleotide sequence, organization and characterization of the atp genes and the encoded subunits of Mycoplasma gallisepticum ATPase.
    Biochem J. 1992 Aug 1;285 ( Pt 3):881-8 PMID: 1386735
  52. Carbon and energy metabolism of atp mutants of Escherichia coli.
    J Bacteriol. 1992 Dec;174(23):7635-41 PMID: 1447134
  53. Organization and sequences of genes for the subunits of ATP synthase in the thermophilic cyanobacterium Synechococcus 6716.
    Biochem J. 1993 Aug 15;294 ( Pt 1):239-51 PMID: 8363578
  54. Location of F1 ATPase-like genes on the physical map of the Bacillus subtilis 168 chromosome.
    Biosci Biotechnol Biochem. 1993 Jul;57(7):1202-3 PMID: 7763993
  55. RocR, a novel regulatory protein controlling arginine utilization in Bacillus subtilis, belongs to the NtrC/NifA family of transcriptional activators.
    J Bacteriol. 1994 Mar;176(5):1234-41 PMID: 8113162
  56. Two compounds implicated in the function of the RC gene of Escherichia coli.
    Nature. 1969 Mar 1;221(5183):838-41 PMID: 4885263
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
Published
1994-11-00
Pages
6802-11
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC197047
Subset
IM
Databases
GENBANK
Z28592
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