Skip to main content
SpringerLink
Log in

The pet genes of Rhodospirillum rubrum: cloning and sequencing of the genes for the cytochrome bc 1-complex

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

A cytochrome bc 1-complex of Rs. rubrum was isolated and the three subunits were purified to homogeneity. The N-terminal amino acid sequence of the purified subunits was determined by automatic Edman degradation. The pet genes of Rhodospirillum rubrum coding for the three subunits of the cytochrome bc 1-complex were isolated from a genomic library of Rs. rubrum using oligonucleotides specific for conserved regions of the subunits from other organisms and a heterologous probe derived from the genes for the complex of Rb. capsulatus. The complete nucleotide sequence of a 5500 by SalI/SphI fragment is described which includes the pet genes and three additional unidentified open reading frames. The N-terminal amino acid sequence of the isolated subunits was used for the identification of the three genes. The genes encoding the subunits are organized as follows: Rieske protein, cytochrome b, cytochrome c 1. Comparison of the N-terminal protein sequences with the protein sequences deduced from the nucleotide sequence showed that only cytochrome c 1 is processed during transport and assembly of the three subunits of the complex. Only the N-terminal methionine of the Rieske protein is cleaved off. The similarity of the deduced amino acid sequence of the three subunits to the corresponding subunits of other organisms is described and implications for structural features of the subunits are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BSA:

bovine serum albumin

SDS:

sodium dodecylsulphate

Rs:

Rhodospirillum

Rb:

Rhodobacter

Pc:

Paracoccus

Rps:

Rhodopseudomonas

References

  • Argos P, Mohana Rao JK, Hargrave PA (1982) Structural prediction of membrane-bound proteins. Eur J Biochem 128:565–575

    Google Scholar 

  • Brasseur R (1988) Calculation of the three-dimensional structure of Saccharomyces cerevisiae cytochrome b inserted in a lipid matrix. J Biol Chem 263:12571–12575

    Google Scholar 

  • Collins MLP, Niederman RA (1976) Membranes of Rhodospirillum rubrum: Physicochemical properties of chromatophore fractions isolated from osmotically and mechanically disrupted cells. J Bacteriol 126:1326–1338

    Google Scholar 

  • Cramer WA, Black MT, Widger WR, Girvan ME (1987) In: Barber J (ed) The light reactions. Elsevier, Amsterdam, pp 447–493

    Google Scholar 

  • Crofts A, Robinson H, Andrews K, Van Doren S, Berry E (1987) Catalytic sites for reduction and oxidation of quinones. In: Papa S, Chance B, Ernster L (eds) Cytochrome systems. Plenum Press, New York, pp 617–624

    Google Scholar 

  • Daldal F, Tokito MK, Davidson E, Faham M (1989) Mutations conferring resistance to quinol oxidation (Qz)-inhibitors of the cytochrome bc1 complex of Rhodobacter capsulatus. EMBO J 8:3951–3961

    Google Scholar 

  • Davidson E, Daldal F (1987) Primary structure of the bc1 complex of Rhodopseudomonas capsulata. Nucleotide sequence of the pet operon encoding the Rieske cytochrome b, and cytochrome c1 apoproteins. J Mol Biol 195:13–24

    Google Scholar 

  • Davis DJ, Frame MK, Johnson DA (1988) Resonance Raman spectroscopy indicates a lysine as the sixth iron ligand in cytochrome f. Biochem Biophys Acta 936:61–66

    Google Scholar 

  • Di Rago JP, Colson AM (1988) Molecular basis for resistance to antimycin and diuron, Q-cycle inhibitors acting at the Qi site in the mitochondrial ubiquinol-cytochrome c reductase in Saccharomyces cerevisiae. J Biol Chem 263:12564–12570

    Google Scholar 

  • Di Rago JP, Coppee JY, Colson AM (1989) Molecular basis for resistance to myxothiazol, mucidin (strobilurin a), and stigmatellin. J Biol Chem 264:14543–14548

    Google Scholar 

  • Eisenberg D, Schwarz E, Komarony M, Wall R (1984) Analysis of membrane and surface protein sequences with the hydrophobic moment plot. J Mol Biol 179:125–142

    Google Scholar 

  • Falk G, Hampe A, Walker E (1985) Nucleotide sequence of the Rhodospirillum rubrum atp operon. Biochem J 228:391–407

    Google Scholar 

  • Gabellini N (1988) Organization and structure of the genes for the cytochrome b/c1 complex in purple photosynthetic bacteria. A phylogenetic study describing the homology of the b/c1, subunits between prokaryotes, mitochondria and chloroplasts. J Bioenerg Biomembr 20:59–83

    Google Scholar 

  • Gabellini N, Harnisch U, McCarthy JEG, Hauska G, Sebald W (1985) 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 4:549–553

    Google Scholar 

  • Gabellini N, Sebald W (1986) Nucleotide sequence and transcription of the fbc operon from Rhodopseudomonas sphaeroides. Eur J Biochem 154:569–579

    Google Scholar 

  • Garger SJ, Griffith OM, Grill LK (1983) Rapid purification of plasmid DNA by single centrifugation in a two-step cesium chloride-ethidium bromide gradient. Biochem Biophys Res Commun 117:835–842

    Google Scholar 

  • Gibson J, Stackebrant E, Zablen LB, Gupta R, Woese CR (1979) A phylogenetic analysis of the purple photosynthetic bacteria. Curr Microbiol 3:59–64

    Google Scholar 

  • Gierasch LM (1989) Signal sequences. Biochemistry 28:923–930

    Google Scholar 

  • Grossberger D (1987) Minipreps of DNA from bacteriophage lambda. Nucleic Acids Res 15:6737

    Google Scholar 

  • Harnisch U, Weiss H, Sebald W (1985) The primary structure of the iron-sulfur subunit of ubiquinol-cytochrome c reductase from Neurospora, determined by cDNA and gene sequencing. Eur J Biochem 149:95–99

    Google Scholar 

  • Hartl FU, Schmidt B, Wachter E, Weiss H, Neupert W (1986) Transport into mitochondria and intramitochondrial sorting of the Fe/S protein of ubiquinol-cytochrome c reductase. Cell 47:939–951

    Google Scholar 

  • Hauska G, HurtE, Gabellini G, Lockau W (1983) Comparative aspects of quinol-cytochrome c/plastocyanin oxidoreductases. Biochem Biophys Acta 726:97–133

    Google Scholar 

  • Hauska G, Nitschke W, Herrmann RG (1988) Amino acid identities in the three redox center carrying polypeptides of cytochrome bc1/b6f-complexes. J Bioenerg Biomembr 20:211–228

    Google Scholar 

  • Henikoff S (1987) Unidirectional digestion with Exonuclease III in DNA sequence analysis. Methods Enzymol 155:156–165

    Google Scholar 

  • Howell N, Gilbert K (1988) Mutational analysis of the mouse mitochondrial cytochrome b gene. Mol Biol 203:607–618

    Google Scholar 

  • Kriauciunas A, Yu L, Yu CA, Wynn RM, Knaff DB (1989) The Rhodospirillum rubrum cytochrome bc1 complex: peptide composition, prosthetic group content and quinone binding. Biochem Biophys Acta 976:70–76

    Google Scholar 

  • Kurowski B, Ludwig B (1987) The genes of the Paracoccus denitrificans bc1 complex. J Biol Chem 262:13805–13811

    Google Scholar 

  • Leustek T, Hartwig R, Weissbach H, Brot N (1988) Regulation of ribulose biphosphate carboxylase expression in Rhodospirillum rubrum: characteristics of mRNA synthesized in vivo and in vitro. J Bacteriol 170:4065–4071

    Google Scholar 

  • Ljungdahl PO, Pennoyer JD, Robertson DE, Trumpower BL (1987) Purification of highly active cytochrome bc1 complexes from phylogenetically diverse species by a single chromatographic procedure. Biochem Biophys Acta 891:227–241

    Google Scholar 

  • Majewski C (1989) The cytochrome bc1 complex of Rhodospirillum rubrum. Isolation of the complex and the coding genes. Thesis, Bochum

  • Malkin R (1988) Structure function studies of photosynthetic cytochrome bc1 and cytochrome b6 complexes. ISI Atlas of Science. Biochemistry: 57–64

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Omerod JG, Omerod KS, Gest H (1961) Light-dependent utilization of organic compounds and photoproduction of molecular hydrogen 18 by photosynthetic bacteria: relationships with nitrogen metabolism. Arch Biochem Biophys 94:449–463

    Google Scholar 

  • Randall LL, Hardy SJS (1989) Unity in function in the absence of consensus in sequence: role of leader peptides in export. Science 243:1156–1159

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    CAS  PubMed  Google Scholar 

  • Saraste M (1984) Localisation of haem-binding sites in the mitochondrial cytochrome b. FEBS Lett 166:367–372

    Google Scholar 

  • Schägger H, Borchart U, Machleidt W, Link TA, Von Jagow G (1987) Isolation and amino acid sequence of the Rieske ironsulfur protein of beef heart ubiquinol cytochrome c reductase. FEBS Lett 219:161–168

    Google Scholar 

  • Silhavy TJ, Berman ML, Enquist LW (1984) Experiments with gene fusions. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Tervoort MJ, Van Gelder BF (1983) The interaction between heme and protein in cytochrome c1. Biochem Biophys Acta 722:137–143

    Google Scholar 

  • Thöny-Meyer L, Stax D, Hennecke H (1989) An unusual gene cluster for the cytochrome bc1 complex in Bradyrhizobium japonicum and its requirement for effective root nodule symbiosis. Cell 57:683–697

    Google Scholar 

  • Van der Wal HN, Van Grondelle R (1983) Flash induced electron transport in b- and c-type cytochromes in Rhodospirillum rubrum. Evidence for a Q-cycle. Biochem Biophys Acta 725:94–103

    Google Scholar 

  • Verbist J, Lang F, Gabellini N, Oesterhelt D (1989) Cloning and sequencing of the fbcF, B and genes encoding the cytochrome b/c1 complex from Rhodopseudomonas viridis. Mol Gen Genet 219:445–452

    Google Scholar 

  • Weiss H, Leonard K, Neupert W (1990) Puzzling subunits of mitochondrial cytochrome reductase. Trends Biochem Sci 15:178–180

    Google Scholar 

  • Widger WR, Cramer WA, Hermann RG, Trebst A (1984) 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 Acta Sci USA 81:674–678

    Google Scholar 

  • Wynn RM, Gaul DF, Choi WK, Shaw RW, Knaff DB (1986) Isolation of cytochrome bc1 complexes from the photosynthetic bacteria Rhodopseudomonas viridis and Rhodospirillum rubrum. Photosyn Res 9:181–195

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Lehrstuhl Biochemie der Pflanzen, Ruhr-Universität, Postfach 10 21 48, W-4630, Bochum 1, Germany

    Christoph Majewski & Achim Trehst

Authors
  1. Christoph Majewski
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Achim Trehst
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by J. Lengeler

The nucleotide sequence reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number X55387

Rights and permissions

Reprints and permissions

About this article

Cite this article

Majewski, C., Trehst, A. The pet genes of Rhodospirillum rubrum: cloning and sequencing of the genes for the cytochrome bc 1-complex. Molec. Gen. Genet. 224, 373–382 (1990). https://doi.org/10.1007/BF00262431

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00262431

Key words

Search

Navigation