Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1986 Jun;166(3):1067–1071. doi: 10.1128/jb.166.3.1067-1071.1986

Nucleotide sequence of the gene for cytochrome b558 of the Bacillus subtilis succinate dehydrogenase complex.

K Magnusson, M K Philips, J R Guest, L Rutberg
PMCID: PMC215233  PMID: 3086287

Abstract

The nucleotide sequence was determined for the first part of the Bacillus subtilis sdh operon. An open reading frame corresponding to the structural gene, sdhA, for cytochrome b558 was identified. The predicted molecular weight of the cytochrome (excluding the N-terminal methionine) is 22,770. It is a very hydrophobic protein with five probable membrane-spanning segments. There is little homology between the B. subtilis cytochrome b558 and cytochrome b of mitochondrial complex III from different organisms or between cytochrome b558 and the hydrophobic sdhC and sdhD peptides of the Escherichia coli sdh operon. About 30 bases downstream of the sdhA stop codon, a new open reading frame starts. The nucleotide sequence predicts the presence of a typical flavin-binding peptide which identifies this reading frame as part of the sdhB gene. Seven bases upstream of the sdhA initiation codon ATG there is a typical B. subtilis ribosome binding site (free energy of interaction, -63 kJ), and further upstream, tentative sigma 55 and sigma 32 promoter sequences were found. The upstream region also contains two 12-base-long direct repeats; their significance is unknown.

Full text

PDF
1067

Selected References

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

  1. Capaldi R. A., Vanderkooi G. The low polarity of many membrane proteins. Proc Natl Acad Sci U S A. 1972 Apr;69(4):930–932. doi: 10.1073/pnas.69.4.930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Condon C., Cammack R., Patil D. S., Owen P. The succinate dehydrogenase of Escherichia coli. Immunochemical resolution and biophysical characterization of a 4-subunit enzyme complex. J Biol Chem. 1985 Aug 5;260(16):9427–9434. [PubMed] [Google Scholar]
  3. Doi R. H. Genetic engineering in Bacillus subtilis. Biotechnol Genet Eng Rev. 1984;2:121–155. doi: 10.1080/02648725.1984.10647797. [DOI] [PubMed] [Google Scholar]
  4. Gabellini N., Harnisch U., McCarthy J. E., Hauska G., Sebald W. Cloning and expression of the fbc operon encoding the FeS protein, cytochrome b and cytochrome c1 from the Rhodopseudomonas sphaeroides b/c1 complex. EMBO J. 1985 Feb;4(2):549–553. doi: 10.1002/j.1460-2075.1985.tb03663.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Green G. N., Kranz J. E., Gennis R. B. Cloning the cyd gene locus coding for the cytochrome d complex of Escherichia coli. Gene. 1984 Dec;32(1-2):99–106. doi: 10.1016/0378-1119(84)90037-4. [DOI] [PubMed] [Google Scholar]
  6. Hasnain S., Sammons R., Roberts I., Thomas C. M. Cloning and deletion analysis of a genomic segment of Bacillus subtilis coding for the sdhA, B, C (succinate dehydrogenase) and gerE (spore germination) loci. J Gen Microbiol. 1985 Sep;131(9):2269–2279. doi: 10.1099/00221287-131-9-2269. [DOI] [PubMed] [Google Scholar]
  7. Hatefi Y. The mitochondrial electron transport and oxidative phosphorylation system. Annu Rev Biochem. 1985;54:1015–1069. doi: 10.1146/annurev.bi.54.070185.005055. [DOI] [PubMed] [Google Scholar]
  8. Hederstedt L. Cytochrome b reducible by succinate in an isolated succinate dehydrogenase-cytochrome b complex from Bacillus subtilis membranes. J Bacteriol. 1980 Dec;144(3):933–940. doi: 10.1128/jb.144.3.933-940.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hederstedt L., Holmgren E., Rutberg L. Characterization of a succinate dehydrogenase complex solubilized from the cytoplasmic membrane of Bacillus subtilis with the nonionic detergent Triton X-100. J Bacteriol. 1979 May;138(2):370–376. doi: 10.1128/jb.138.2.370-376.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hederstedt L., Rutberg L. Orientation of succinate dehydrogenase and cytochrome b558 in the Bacillus subtilis cytoplasmic membrane. J Bacteriol. 1983 Jan;153(1):57–65. doi: 10.1128/jb.153.1.57-65.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hederstedt L., Rutberg L. Succinate dehydrogenase--a comparative review. Microbiol Rev. 1981 Dec;45(4):542–555. doi: 10.1128/mr.45.4.542-555.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnson W. C., Moran C. P., Jr, Losick R. Two RNA polymerase sigma factors from Bacillus subtilis discriminate between overlapping promoters for a developmentally regulated gene. Nature. 1983 Apr 28;302(5911):800–804. doi: 10.1038/302800a0. [DOI] [PubMed] [Google Scholar]
  13. Kabsch W., Sander C. On the use of sequence homologies to predict protein structure: identical pentapeptides can have completely different conformations. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1075–1078. doi: 10.1073/pnas.81.4.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kita K., Yamato I., Anraku Y. Purification and properties of cytochrome b556 in the respiratory chain of aerobically grown Escherichia coli K12. J Biol Chem. 1978 Dec 25;253(24):8910–8915. [PubMed] [Google Scholar]
  15. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  16. Magnusson K., Hederstedt L., Rutberg L. Cloning and expression in Escherichia coli of sdhA, the structural gene for cytochrome b558 of the Bacillus subtilis succinate dehydrogenase complex. J Bacteriol. 1985 Jun;162(3):1180–1185. doi: 10.1128/jb.162.3.1180-1185.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Magnusson K., Rutberg B., Hederstedt L., Rutberg L. Characterization of a pleiotropic succinate dehydrogenase-negative mutant of Bacillus subtilis. J Gen Microbiol. 1983 Apr;129(4):917–922. doi: 10.1099/00221287-129-4-917. [DOI] [PubMed] [Google Scholar]
  18. Messing J., Crea R., Seeburg P. H. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. doi: 10.1093/nar/9.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miles J. S., Guest J. R. Complete nucleotide sequence of the fumarase gene (citG) of Bacillus subtilis 168. Nucleic Acids Res. 1985 Jan 11;13(1):131–140. doi: 10.1093/nar/13.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  21. Murakami H., Kita K., Anraku Y. Cloning of cybB, the gene for cytochrome b561 of Escherichia coli K12. Mol Gen Genet. 1984;198(2):1–6. doi: 10.1007/BF00328692. [DOI] [PubMed] [Google Scholar]
  22. Murakami H., Kita K., Oya H., Anraku Y. Chromosomal location of the Escherichia coli cytochrome b556 gene, cybA. Mol Gen Genet. 1984;196(1):1–5. doi: 10.1007/BF00334084. [DOI] [PubMed] [Google Scholar]
  23. Ohné M. Regulation of the dicarboxylic acid part of the citric acid cycle in Bacillus subtilis. J Bacteriol. 1975 Apr;122(1):224–234. doi: 10.1128/jb.122.1.224-234.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Widger W. R., Cramer W. A., Herrmann R. G., Trebst A. Sequence homology and structural similarity between cytochrome b of mitochondrial complex III and the chloroplast b6-f complex: position of the cytochrome b hemes in the membrane. Proc Natl Acad Sci U S A. 1984 Feb;81(3):674–678. doi: 10.1073/pnas.81.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wong S. L., Price C. W., Goldfarb D. S., Doi R. H. The subtilisin E gene of Bacillus subtilis is transcribed from a sigma 37 promoter in vivo. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1184–1188. doi: 10.1073/pnas.81.4.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wood D., Darlison M. G., Wilde R. J., Guest J. R. Nucleotide sequence encoding the flavoprotein and hydrophobic subunits of the succinate dehydrogenase of Escherichia coli. Biochem J. 1984 Sep 1;222(2):519–534. doi: 10.1042/bj2220519. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES