Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1993 Apr;12(4):1283–1292. doi: 10.1002/j.1460-2075.1993.tb05773.x

A novel membrane-associated c-type cytochrome, cyt cy, can mediate the photosynthetic growth of Rhodobacter capsulatus and Rhodobacter sphaeroides.

F E Jenney Jr 1, F Daldal 1
PMCID: PMC413338  PMID: 8385603

Abstract

Mutants of Rhodobacter capsulatus lacking the soluble electron carrier cytochrome c2 are able to grow photosynthetically (Ps+), whereas Rhodobacter sphaeroides is unable to do so. To understand this unusual electron transfer pathway the gene required for cyt c2-independent growth of R.capsulatus was sought using chromosomal libraries derived from a cyt c2- mutant of this species to complement a Ps- cyt c2- mutant of R.sphaeroides to Ps+ growth. The complementing 1.2 kbp DNA fragment contained a gene, cycY, encoding a novel membrane-associated c-type cytochrome, cyt cy, based on predicted amino acid sequence, optical difference spectra and SDS-PAGE analysis of chromatophore membranes. The predicted primary sequence of cyt cy is unusual in having two distinct domains, a hydrophobic amino-terminal region and a carboxyl-terminus with strong homology to cytochromes c. A cyt cy- mutant of R.capsulatus remains Ps+ as does the cyt c2- mutant. However, a mutant lacking both cyt c2 and cy is Ps-, and can be complemented to Ps+ by either cyt c2 or cyt cy. These findings demonstrate that each of the cytochromes c2 and cy is essential for photosynthesis only in the absence of the other. Thus, two distinct electron transfer pathways, unrecognized until now, operate during photosynthesis in R.capsulatus under appropriate conditions, one via the soluble cyt c2 and the other via the membrane-associated cyt cy.

Full text

PDF
1283

Images in this article

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Atta-Asafo-Adjei E., Daldal F. Size of the amino acid side chain at position 158 of cytochrome b is critical for an active cytochrome bc1 complex and for photosynthetic growth of Rhodobacter capsulatus. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):492–496. doi: 10.1073/pnas.88.2.492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benning M. M., Wesenberg G., Caffrey M. S., Bartsch R. G., Meyer T. E., Cusanovich M. A., Rayment I., Holden H. M. Molecular structure of cytochrome c2 isolated from Rhodobacter capsulatus determined at 2.5 A resolution. J Mol Biol. 1991 Aug 5;220(3):673–685. doi: 10.1016/0022-2836(91)90109-j. [DOI] [PubMed] [Google Scholar]
  4. Berry E. A., Trumpower B. L. Isolation of ubiquinol oxidase from Paracoccus denitrificans and resolution into cytochrome bc1 and cytochrome c-aa3 complexes. J Biol Chem. 1985 Feb 25;260(4):2458–2467. [PubMed] [Google Scholar]
  5. Bott M., Ritz D., Hennecke H. The Bradyrhizobium japonicum cycM gene encodes a membrane-anchored homolog of mitochondrial cytochrome c. J Bacteriol. 1991 Nov;173(21):6766–6772. doi: 10.1128/jb.173.21.6766-6772.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caffrey M., Davidson E., Cusanovich M., Daldal F. Cytochrome c2 mutants of Rhodobacter capsulatus. Arch Biochem Biophys. 1992 Feb 1;292(2):419–426. doi: 10.1016/0003-9861(92)90011-k. [DOI] [PubMed] [Google Scholar]
  7. Daldal F., Cheng S., Applebaum J., Davidson E., Prince R. C. Cytochrome c(2) is not essential for photosynthetic growth of Rhodopseudomonas capsulata. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2012–2016. doi: 10.1073/pnas.83.7.2012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davidson E., Prince R. C., Haith C. E., Daldal F. The cytochrome bc1 complex of Rhodobacter sphaeroides can restore cytochrome c2-independent photosynthetic growth to a Rhodobacter capsulatus mutant lacking cytochrome bc1. J Bacteriol. 1989 Nov;171(11):6059–6068. doi: 10.1128/jb.171.11.6059-6068.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deisenhofer J., Michel H. The Photosynthetic Reaction Center from the Purple Bacterium Rhodopseudomonas viridis. Science. 1989 Sep 29;245(4925):1463–1473. doi: 10.1126/science.245.4925.1463. [DOI] [PubMed] [Google Scholar]
  10. Ditta G., Schmidhauser T., Yakobson E., Lu P., Liang X. W., Finlay D. R., Guiney D., Helinski D. R. Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid. 1985 Mar;13(2):149–153. doi: 10.1016/0147-619x(85)90068-x. [DOI] [PubMed] [Google Scholar]
  11. Donohue T. J., Kaplan S. Genetic techniques in rhodospirillaceae. Methods Enzymol. 1991;204:459–485. doi: 10.1016/0076-6879(91)04024-i. [DOI] [PubMed] [Google Scholar]
  12. Donohue T. J., McEwan A. G., Van Doren S., Crofts A. R., Kaplan S. Phenotypic and genetic characterization of cytochrome c2 deficient mutants of Rhodobacter sphaeroides. Biochemistry. 1988 Mar 22;27(6):1918–1925. doi: 10.1021/bi00406a018. [DOI] [PubMed] [Google Scholar]
  13. Fitch J., Cannac V., Meyer T. E., Cusanovich M. A., Tollin G., Van Beeumen J., Rott M. A., Donohue T. J. Expression of a cytochrome c2 isozyme restores photosynthetic growth of Rhodobacter sphaeroides mutants lacking the wild-type cytochrome c2 gene. Arch Biochem Biophys. 1989 Jun;271(2):502–507. doi: 10.1016/0003-9861(89)90301-9. [DOI] [PubMed] [Google Scholar]
  14. Jones M. R., McEwan A. G., Jackson J. B. The role of c-type cytochromes in the photosynthetic electron transport pathway of Rhodobacter capsulatus. Biochim Biophys Acta. 1990 Aug 9;1019(1):59–66. doi: 10.1016/0005-2728(90)90124-m. [DOI] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Marrs B., Gest H. Genetic mutations affecting the respiratory electron-transport system of the photosynthetic bacterium Rhodopseudomonas capsulata. J Bacteriol. 1973 Jun;114(3):1045–1051. doi: 10.1128/jb.114.3.1045-1051.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Melandri A. B., Zannoni D. Photosynthetic and respiratory electron flow in the dual functional membrane of facultative photosynthetic bacteria. J Bioenerg Biomembr. 1978 Aug;10(3-4):109–138. doi: 10.1007/BF00743056. [DOI] [PubMed] [Google Scholar]
  18. Okkels J. S., Kjaer B., Hansson O., Svendsen I., Møller B. L., Scheller H. V. A membrane-bound monoheme cytochrome c551 of a novel type is the immediate electron donor to P840 of the Chlorobium vibrioforme photosynthetic reaction center complex. J Biol Chem. 1992 Oct 15;267(29):21139–21145. [PubMed] [Google Scholar]
  19. Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
  20. Prince R. C., Daldal F. Physiological electron donors to the photochemical reaction center of Rhodobacter capsulatus. Biochim Biophys Acta. 1987 Dec 17;894(3):370–378. doi: 10.1016/0005-2728(87)90115-0. [DOI] [PubMed] [Google Scholar]
  21. Robertson D. E., Dutton P. L. The nature and magnitude of the charge-separation reactions of ubiquinol cytochrome c2 oxidoreductase. Biochim Biophys Acta. 1988 Oct 5;935(3):273–291. doi: 10.1016/0005-2728(88)90223-x. [DOI] [PubMed] [Google Scholar]
  22. SISTROM W. R. A requirement for sodium in the growth of Rhodopseudomonas spheroides. J Gen Microbiol. 1960 Jun;22:778–785. doi: 10.1099/00221287-22-3-778. [DOI] [PubMed] [Google Scholar]
  23. Scolnik P. A., Marrs B. L. Genetic research with photosynthetic bacteria. Annu Rev Microbiol. 1987;41:703–726. doi: 10.1146/annurev.mi.41.100187.003415. [DOI] [PubMed] [Google Scholar]
  24. Sone N., Sekimachi M., Kutoh E. Identification and properties of a quinol oxidase super-complex composed of a bc1 complex and cytochrome oxidase in the thermophilic bacterium PS3. J Biol Chem. 1987 Nov 15;262(32):15386–15391. [PubMed] [Google Scholar]
  25. Thomas P. E., Ryan D., Levin W. An improved staining procedure for the detection of the peroxidase activity of cytochrome P-450 on sodium dodecyl sulfate polyacrylamide gels. Anal Biochem. 1976 Sep;75(1):168–176. doi: 10.1016/0003-2697(76)90067-1. [DOI] [PubMed] [Google Scholar]
  26. Venturoli G., Gabellini N., Oesterhelt D., Melandri B. A. Kinetics of photosynthetic electron transfer in artificial vesicles reconstituted with purified complexes from Rhodobacter capsulatus. II. Direct electron transfer between the reaction center and the bc1 complex and role of cytochrome c2. Eur J Biochem. 1990 Apr 20;189(1):95–103. doi: 10.1111/j.1432-1033.1990.tb15464.x. [DOI] [PubMed] [Google Scholar]
  27. Yen H. C., Hu N. T., Marrs B. L. Characterization of the gene transfer agent made by an overproducer mutant of Rhodopseudomonas capsulata. J Mol Biol. 1979 Jun 25;131(2):157–168. doi: 10.1016/0022-2836(79)90071-8. [DOI] [PubMed] [Google Scholar]
  28. von Heijne G. Transcending the impenetrable: how proteins come to terms with membranes. Biochim Biophys Acta. 1988 Jun 9;947(2):307–333. doi: 10.1016/0304-4157(88)90013-5. [DOI] [PubMed] [Google Scholar]
  29. von Wachenfeldt C., Hederstedt L. Bacillus subtilis 13-kilodalton cytochrome c-550 encoded by cccA consists of a membrane-anchor and a heme domain. J Biol Chem. 1990 Aug 15;265(23):13939–13948. [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES