Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1990 Aug;172(8):4497–4504. doi: 10.1128/jb.172.8.4497-4504.1990

Gene encoding the 5.7-kilodalton chlorosome protein of Chloroflexus aurantiacus: regulated message levels and a predicted carboxy-terminal protein extension.

S J Theroux 1, T E Redlinger 1, R C Fuller 1, S J Robinson 1
PMCID: PMC213280  PMID: 2376566

Abstract

The major light-harvesting pigment of the green filamentous bacterium Chloroflexus aurantiacus is bacteriochlorophyll (Bchl) c, localized in chlorosomes attached to the inner surface of the cytoplasmic membrane. Chlorosomes consist of four polypeptides and associated pigments and lipids. Previous studies of the inducible assembly of the photosynthetic apparatus had indicated that the major chlorosomal polypeptides are present as high-molecular-weight aggregates before the appearance of mature chlorosomes, and a mechanism for posttranslational processing of a polyprotein had been proposed. We have isolated the gene (csmA) encoding the 5.7-kilodalton chlorosomal polypeptide from C. aurantiacus in order to determine whether this protein is synthesized as part of a polyprotein. Analysis of the nucleotide sequence of csmA indicates that the gene is not large enough to encode more than one known chlorosome polypeptide. Transcriptional analysis indicates that csmA is transcribed as a small message whose abundance is regulated in response to oxygen, so that no csmA message is detectable in cells grown aerobically in the dark. Comparison of the sequence predicted by csmA with the peptide sequence of the Bchl c binding protein purified from chlorosomes indicates that this protein is synthesized with a carboxy-terminal extension of 27 amino acids. We discuss possible roles for this carboxy-terminal extension in the assembly of chlorosomes.

Full text

PDF
4497

Images in this article

4498Image
on p.4498
4500Image
on p.4500
4501Image
on p.4501
4501Image
on p.4501
4502Image
on p.4502

Selected References

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

  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. 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]
  3. Belasco J. G., Beatty J. T., Adams C. W., von Gabain A., Cohen S. N. Differential expression of photosynthesis genes in R. capsulata results from segmental differences in stability within the polycistronic rxcA transcript. Cell. 1985 Jan;40(1):171–181. doi: 10.1016/0092-8674(85)90320-4. [DOI] [PubMed] [Google Scholar]
  4. Birkmann A., Sawers R. G., Böck A. Involvement of the ntrA gene product in the anaerobic metabolism of Escherichia coli. Mol Gen Genet. 1987 Dec;210(3):535–542. doi: 10.1007/BF00327209. [DOI] [PubMed] [Google Scholar]
  5. Brune D. C., Nozawa T., Blankenship R. E. Antenna organization in green photosynthetic bacteria. 1. Oligomeric bacteriochlorophyll c as a model for the 740 nm absorbing bacteriochlorophyll c in Chloroflexus aurantiacus chlorosomes. Biochemistry. 1987 Dec 29;26(26):8644–8652. doi: 10.1021/bi00400a023. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Chen C. Y., Beatty J. T., Cohen S. N., Belasco J. G. An intercistronic stem-loop structure functions as an mRNA decay terminator necessary but insufficient for puf mRNA stability. Cell. 1988 Feb 26;52(4):609–619. doi: 10.1016/0092-8674(88)90473-4. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. David M., Daveran M. L., Batut J., Dedieu A., Domergue O., Ghai J., Hertig C., Boistard P., Kahn D. Cascade regulation of nif gene expression in Rhizobium meliloti. Cell. 1988 Aug 26;54(5):671–683. doi: 10.1016/s0092-8674(88)80012-6. [DOI] [PubMed] [Google Scholar]
  10. DeHoff B. S., Lee J. K., Donohue T. J., Gumport R. I., Kaplan S. In vivo analysis of puf operon expression in Rhodobacter sphaeroides after deletion of a putative intercistronic transcription terminator. J Bacteriol. 1988 Oct;170(10):4681–4692. doi: 10.1128/jb.170.10.4681-4692.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Diner B. A., Ries D. F., Cohen B. N., Metz J. G. COOH-terminal processing of polypeptide D1 of the photosystem II reaction center of Scenedesmus obliquus is necessary for the assembly of the oxygen-evolving complex. J Biol Chem. 1988 Jun 25;263(18):8972–8980. [PubMed] [Google Scholar]
  13. Drews G., Oelze J. Organization and differentiation of membranes of phototrophic bacteria. Adv Microb Physiol. 1981;22:1–92. doi: 10.1016/s0065-2911(08)60325-2. [DOI] [PubMed] [Google Scholar]
  14. Dreyfus M. What constitutes the signal for the initiation of protein synthesis on Escherichia coli mRNAs? J Mol Biol. 1988 Nov 5;204(1):79–94. doi: 10.1016/0022-2836(88)90601-8. [DOI] [PubMed] [Google Scholar]
  15. Ellis J. Proteins as molecular chaperones. 1987 Jul 30-Aug 5Nature. 328(6129):378–379. doi: 10.1038/328378a0. [DOI] [PubMed] [Google Scholar]
  16. Feick R. G., Fitzpatrick M., Fuller R. C. Isolation and characterization of cytoplasmic membranes and chlorosomes from the green bacterium Chloroflexus aurantiacus. J Bacteriol. 1982 May;150(2):905–915. doi: 10.1128/jb.150.2.905-915.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Finley D., Bartel B., Varshavsky A. The tails of ubiquitin precursors are ribosomal proteins whose fusion to ubiquitin facilitates ribosome biogenesis. Nature. 1989 Mar 30;338(6214):394–401. doi: 10.1038/338394a0. [DOI] [PubMed] [Google Scholar]
  18. Foster J. M., Redlinger T. E., Blankenship R. E., Fuller R. C. Oxygen regulation of development of the photosynthetic membrane system in Chloroflexus aurantiacus. J Bacteriol. 1986 Aug;167(2):655–659. doi: 10.1128/jb.167.2.655-659.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gould S. G., Keller G. A., Subramani S. Identification of a peroxisomal targeting signal at the carboxy terminus of firefly luciferase. J Cell Biol. 1987 Dec;105(6 Pt 2):2923–2931. doi: 10.1083/jcb.105.6.2923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gussin G. N., Ronson C. W., Ausubel F. M. Regulation of nitrogen fixation genes. Annu Rev Genet. 1986;20:567–591. doi: 10.1146/annurev.ge.20.120186.003031. [DOI] [PubMed] [Google Scholar]
  21. Hanahan D., Meselson M. Plasmid screening at high colony density. Methods Enzymol. 1983;100:333–342. doi: 10.1016/0076-6879(83)00066-x. [DOI] [PubMed] [Google Scholar]
  22. Hemmingsen S. M., Woolford C., van der Vies S. M., Tilly K., Dennis D. T., Georgopoulos C. P., Hendrix R. W., Ellis R. J. Homologous plant and bacterial proteins chaperone oligomeric protein assembly. Nature. 1988 May 26;333(6171):330–334. doi: 10.1038/333330a0. [DOI] [PubMed] [Google Scholar]
  23. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  24. Kassavetis G. A., Geiduschek E. P. Bacteriophage T4 late promoters: mapping 5' ends of T4 gene 23 mRNAs. EMBO J. 1982;1(1):107–114. doi: 10.1002/j.1460-2075.1982.tb01132.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kiley P. J., Kaplan S. Molecular genetics of photosynthetic membrane biosynthesis in Rhodobacter sphaeroides. Microbiol Rev. 1988 Mar;52(1):50–69. doi: 10.1128/mr.52.1.50-69.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kustu S., Santero E., Keener J., Popham D., Weiss D. Expression of sigma 54 (ntrA)-dependent genes is probably united by a common mechanism. Microbiol Rev. 1989 Sep;53(3):367–376. doi: 10.1128/mr.53.3.367-376.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mains R. E., Eipper B. A., Ling N. Common precursor to corticotropins and endorphins. Proc Natl Acad Sci U S A. 1977 Jul;74(7):3014–3018. doi: 10.1073/pnas.74.7.3014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  30. Nixon B. T., Ronson C. W., Ausubel F. M. Two-component regulatory systems responsive to environmental stimuli share strongly conserved domains with the nitrogen assimilation regulatory genes ntrB and ntrC. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7850–7854. doi: 10.1073/pnas.83.20.7850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ovchinnikov YuA, Abdulaev N. G., Zolotarev A. S., Shmukler B. E., Zargarov A. A., Kutuzov M. A., Telezhinskaya I. N., Levina N. B. Photosynthetic reaction centre of Chloroflexus aurantiacus. I. Primary structure of L-subunit. FEBS Lett. 1988 Apr 11;231(1):237–242. doi: 10.1016/0014-5793(88)80739-7. [DOI] [PubMed] [Google Scholar]
  32. Oyaizu H., Debrunner-Vossbrinck B., Mandelco L., Studier J. A., Woese C. R. The green non-sulfur bacteria: a deep branching in the eubacterial line of descent. Syst Appl Microbiol. 1987;9:47–53. doi: 10.1016/s0723-2020(87)80055-3. [DOI] [PubMed] [Google Scholar]
  33. Ronson C. W., Nixon B. T., Albright L. M., Ausubel F. M. Rhizobium meliloti ntrA (rpoN) gene is required for diverse metabolic functions. J Bacteriol. 1987 Jun;169(6):2424–2431. doi: 10.1128/jb.169.6.2424-2431.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Smith D. R., Calvo J. M. Nucleotide sequence of the E coli gene coding for dihydrofolate reductase. Nucleic Acids Res. 1980 May 24;8(10):2255–2274. doi: 10.1093/nar/8.10.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sprague S. G., Staehelin L. A., DiBartolomeis M. J., Fuller R. C. Isolation and development of chlorosomes in the green bacterium Chloroflexus aurantiacus. J Bacteriol. 1981 Sep;147(3):1021–1031. doi: 10.1128/jb.147.3.1021-1031.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sprague S. G., Staehelin L. A., Fuller R. C. Semiaerobic induction of bacteriochlorophyll synthesis in the green bacterium Chloroflexus aurantiacus. J Bacteriol. 1981 Sep;147(3):1032–1039. doi: 10.1128/jb.147.3.1032-1039.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Stock J. B., Ninfa A. J., Stock A. M. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev. 1989 Dec;53(4):450–490. doi: 10.1128/mr.53.4.450-490.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Swinkels B. W., Evers R., Borst P. The topogenic signal of the glycosomal (microbody) phosphoglycerate kinase of Crithidia fasciculata resides in a carboxy-terminal extension. EMBO J. 1988 Apr;7(4):1159–1165. doi: 10.1002/j.1460-2075.1988.tb02926.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Verner K., Schatz G. Protein translocation across membranes. Science. 1988 Sep 9;241(4871):1307–1313. doi: 10.1126/science.2842866. [DOI] [PubMed] [Google Scholar]
  43. Yang R., Lis J., Wu R. Elution of DNA from agarose gels after electrophoresis. Methods Enzymol. 1979;68:176–182. doi: 10.1016/0076-6879(79)68012-6. [DOI] [PubMed] [Google Scholar]

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

RESOURCES