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. 1995 Jun;177(11):2990–2997. doi: 10.1128/jb.177.11.2990-2997.1995

Characterization of an aerobic repressor that coordinately regulates bacteriochlorophyll, carotenoid, and light harvesting-II expression in Rhodobacter capsulatus.

S N Ponnampalam 1, J J Buggy 1, C E Bauer 1
PMCID: PMC176984  PMID: 7768793

Abstract

For most species of purple photosynthetic bacteria, the presence of molecular oxygen represses synthesis of carotenoids and bacteriochlorophyll. In this study we characterize a strain of Rhodobacter capsulatus, DB469, which contains a genomic disruption of an open reading frame in the photosynthesis gene cluster termed ORF469. Characterization of the steady-state level of bacteriochlorophyll synthesis demonstrates that disruption of ORF469 results in a 2.5-fold increase in aerobic synthesis of bacteriochlorophyll over that observed with the parent strain. Utilizing reporter plasmids that contain transcriptional fusions of lacZ to various carotenoid and bacteriochlorophyll biosynthesis genes, we also demonstrate that disruption of ORF469 leads to an approximate twofold increase in bacteriochlorophyll and carotenoid gene expression under anaerobic growth conditions. Similar analysis with reporter plasmids that contain translational fusions of lacZ to the puf, puh, and puc operons demonstrates that disruption of ORF469 leads to elevated levels of aerobic transcription of light harvesting-II genes (puc), without affecting light harvesting-I or reaction center gene expression (puf and puh, respectively). Gel mobility analysis demonstrates that DB469 cells lack a DNA-binding protein that interacts with a palindromic sequence in the bchC promoter region. The results of this study indicate that ORF469 codes for a DNA-binding protein that acts as an aerobic repressor of promoters for bacteriochlorophyll, carotenoid, and light harvesting-II gene expression.

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Selected References

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  1. Adams C. W., Forrest M. E., Cohen S. N., Beatty J. T. Structural and functional analysis of transcriptional control of the Rhodobacter capsulatus puf operon. J Bacteriol. 1989 Jan;171(1):473–482. doi: 10.1128/jb.171.1.473-482.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Armstrong G. A., Alberti M., Leach F., Hearst J. E. Nucleotide sequence, organization, and nature of the protein products of the carotenoid biosynthesis gene cluster of Rhodobacter capsulatus. Mol Gen Genet. 1989 Apr;216(2-3):254–268. doi: 10.1007/BF00334364. [DOI] [PubMed] [Google Scholar]
  3. Bauer C. E., Buggy J. J., Yang Z. M., Marrs B. L. The superoperonal organization of genes for pigment biosynthesis and reaction center proteins is a conserved feature in Rhodobacter capsulatus: analysis of overlapping bchB and puhA transcripts. Mol Gen Genet. 1991 Sep;228(3):433–444. doi: 10.1007/BF00260637. [DOI] [PubMed] [Google Scholar]
  4. Bauer C. E., Young D. A., Marrs B. L. Analysis of the Rhodobacter capsulatus puf operon. Location of the oxygen-regulated promoter region and the identification of an additional puf-encoded gene. J Biol Chem. 1988 Apr 5;263(10):4820–4827. [PubMed] [Google Scholar]
  5. Bauer C., Buggy J., Mosley C. Control of photosystem genes in Rhodobacter capsulatus. Trends Genet. 1993 Feb;9(2):56–60. doi: 10.1016/0168-9525(93)90188-N. [DOI] [PubMed] [Google Scholar]
  6. Biel A. J., Marrs B. L. Transcriptional regulation of several genes for bacteriochlorophyll biosynthesis in Rhodopseudomonas capsulata in response to oxygen. J Bacteriol. 1983 Nov;156(2):686–694. doi: 10.1128/jb.156.2.686-694.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bollivar D. W., Suzuki J. Y., Beatty J. T., Dobrowolski J. M., Bauer C. E. Directed mutational analysis of bacteriochlorophyll a biosynthesis in Rhodobacter capsulatus. J Mol Biol. 1994 Apr 15;237(5):622–640. doi: 10.1006/jmbi.1994.1260. [DOI] [PubMed] [Google Scholar]
  8. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  9. Buggy J. J., Sganga M. W., Bauer C. E. Characterization of a light-responding trans-activator responsible for differentially controlling reaction center and light-harvesting-I gene expression in Rhodobacter capsulatus. J Bacteriol. 1994 Nov;176(22):6936–6943. doi: 10.1128/jb.176.22.6936-6943.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. COHEN-BAZIRE G., SISTROM W. R., STANIER R. Y. Kinetic studies of pigment synthesis by non-sulfur purple bacteria. J Cell Physiol. 1957 Feb;49(1):25–68. doi: 10.1002/jcp.1030490104. [DOI] [PubMed] [Google Scholar]
  11. Clark W. G., Davidson E., Marrs B. L. Variation of levels of mRNA coding for antenna and reaction center polypeptides in Rhodopseudomonas capsulata in response to changes in oxygen concentration. J Bacteriol. 1984 Mar;157(3):945–948. doi: 10.1128/jb.157.3.945-948.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Donohue T. J., Hoger J. H., Kaplan S. Cloning and expression of the Rhodobacter sphaeroides reaction center H gene. J Bacteriol. 1986 Nov;168(2):953–961. doi: 10.1128/jb.168.2.953-961.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fonstein M., Haselkorn R. Chromosomal structure of Rhodobacter capsulatus strain SB1003: cosmid encyclopedia and high-resolution physical and genetic map. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2522–2526. doi: 10.1073/pnas.90.6.2522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gough J. A., Murray N. E. Sequence diversity among related genes for recognition of specific targets in DNA molecules. J Mol Biol. 1983 May 5;166(1):1–19. doi: 10.1016/s0022-2836(83)80047-3. [DOI] [PubMed] [Google Scholar]
  15. Harrison S. C., Aggarwal A. K. DNA recognition by proteins with the helix-turn-helix motif. Annu Rev Biochem. 1990;59:933–969. doi: 10.1146/annurev.bi.59.070190.004441. [DOI] [PubMed] [Google Scholar]
  16. Hunter C. N., McGlynn P., Ashby M. K., Burgess J. G., Olsen J. D. DNA sequencing and complementation/deletion analysis of the bchA-puf operon region of Rhodobacter sphaeroides: in vivo mapping of the oxygen-regulated puf promoter. Mol Microbiol. 1991 Nov;5(11):2649–2661. doi: 10.1111/j.1365-2958.1991.tb01974.x. [DOI] [PubMed] [Google Scholar]
  17. Inoue K., Kouadio J. L., Mosley C. S., Bauer C. E. Isolation and in vitro phosphorylation of sensory transduction components controlling anaerobic induction of light harvesting and reaction center gene expression in Rhodobacter capsulatus. Biochemistry. 1995 Jan 17;34(2):391–396. doi: 10.1021/bi00002a002. [DOI] [PubMed] [Google Scholar]
  18. Klug G., Kaufmann N., Drews G. Gene expression of pigment-binding proteins of the bacterial photosynthetic apparatus: Transcription and assembly in the membrane of Rhodopseudomonas capsulata. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6485–6489. doi: 10.1073/pnas.82.19.6485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lee J. K., Kaplan S. cis-acting regulatory elements involved in oxygen and light control of puc operon transcription in Rhodobacter sphaeroides. J Bacteriol. 1992 Feb;174(4):1146–1157. doi: 10.1128/jb.174.4.1146-1157.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ma D., Cook D. N., O'Brien D. A., Hearst J. E. Analysis of the promoter and regulatory sequences of an oxygen-regulated bch operon in Rhodobacter capsulatus by site-directed mutagenesis. J Bacteriol. 1993 Apr;175(7):2037–2045. doi: 10.1128/jb.175.7.2037-2045.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Marrs B. Mobilization of the genes for photosynthesis from Rhodopseudomonas capsulata by a promiscuous plasmid. J Bacteriol. 1981 Jun;146(3):1003–1012. doi: 10.1128/jb.146.3.1003-1012.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McGlynn P., Hunter C. N. Genetic analysis of the bchC and bchA genes of Rhodobacter sphaeroides. Mol Gen Genet. 1993 Jan;236(2-3):227–234. doi: 10.1007/BF00277117. [DOI] [PubMed] [Google Scholar]
  23. Minton N. P. Improved plasmid vectors for the isolation of translational lac gene fusions. Gene. 1984 Nov;31(1-3):269–273. doi: 10.1016/0378-1119(84)90220-8. [DOI] [PubMed] [Google Scholar]
  24. Mosley C. S., Suzuki J. Y., Bauer C. E. Identification and molecular genetic characterization of a sensor kinase responsible for coordinately regulating light harvesting and reaction center gene expression in response to anaerobiosis. J Bacteriol. 1994 Dec;176(24):7566–7573. doi: 10.1128/jb.176.24.7566-7573.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Penfold R. J., Pemberton J. M. Sequencing, chromosomal inactivation, and functional expression in Escherichia coli of ppsR, a gene which represses carotenoid and bacteriochlorophyll synthesis in Rhodobacter sphaeroides. J Bacteriol. 1994 May;176(10):2869–2876. doi: 10.1128/jb.176.10.2869-2876.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Sganga M. W., Bauer C. E. Regulatory factors controlling photosynthetic reaction center and light-harvesting gene expression in Rhodobacter capsulatus. Cell. 1992 Mar 6;68(5):945–954. doi: 10.1016/0092-8674(92)90037-d. [DOI] [PubMed] [Google Scholar]
  28. Taylor D. P., Cohen S. N., Clark W. G., Marrs B. L. Alignment of genetic and restriction maps of the photosynthesis region of the Rhodopseudomonas capsulata chromosome by a conjugation-mediated marker rescue technique. J Bacteriol. 1983 May;154(2):580–590. doi: 10.1128/jb.154.2.580-590.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tichy H. V., Albien K. U., Gad'on N., Drews G. Analysis of the Rhodobacter capsulatus puc operon: the pucC gene plays a central role in the regulation of LHII (B800-850 complex) expression. EMBO J. 1991 Oct;10(10):2949–2955. doi: 10.1002/j.1460-2075.1991.tb07845.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wellington C. L., Beatty J. T. Promoter mapping and nucleotide sequence of the bchC bacteriochlorophyll biosynthesis gene from Rhodobacter capsulatus. Gene. 1989 Nov 30;83(2):251–261. doi: 10.1016/0378-1119(89)90111-x. [DOI] [PubMed] [Google Scholar]
  31. Yen H. C., Marrs B. Map of genes for carotenoid and bacteriochlorophyll biosynthesis in Rhodopseudomonas capsulata. J Bacteriol. 1976 May;126(2):619–629. doi: 10.1128/jb.126.2.619-629.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Young D. A., Bauer C. E., Williams J. C., Marrs B. L. Genetic evidence for superoperonal organization of genes for photosynthetic pigments and pigment-binding proteins in Rhodobacter capsulatus. Mol Gen Genet. 1989 Jul;218(1):1–12. doi: 10.1007/BF00330558. [DOI] [PubMed] [Google Scholar]
  33. Youvan D. C., Ismail S. Light-harvesting II (B800-B850 complex) structural genes from Rhodopseudomonas capsulata. Proc Natl Acad Sci U S A. 1985 Jan;82(1):58–62. doi: 10.1073/pnas.82.1.58. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zhu Y. S., Hearst J. E. Regulation of expression of genes for light-harvesting antenna proteins LH-I and LH-II; reaction center polypeptides RC-L, RC-M, and RC-H; and enzymes of bacteriochlorophyll and carotenoid biosynthesis in Rhodobacter capsulatus by light and oxygen. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7613–7617. doi: 10.1073/pnas.83.20.7613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zsebo K. M., Hearst J. E. Genetic-physical mapping of a photosynthetic gene cluster from R. capsulata. Cell. 1984 Jul;37(3):937–947. doi: 10.1016/0092-8674(84)90428-8. [DOI] [PubMed] [Google Scholar]
  36. Zucconi A. P., Beatty J. T. Posttranscriptional regulation by light of the steady-state levels of mature B800-850 light-harvesting complexes in Rhodobacter capsulatus. J Bacteriol. 1988 Feb;170(2):877–882. doi: 10.1128/jb.170.2.877-882.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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