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. 1989 May;8(5):1303–1308. doi: 10.1002/j.1460-2075.1989.tb03509.x

Multiple copies of the coding regions for the light-harvesting B800-850 alpha- and beta-polypeptides are present in the Rhodopseudomonas palustris genome.

M H Tadros 1, K Waterkamp 1
PMCID: PMC400955  PMID: 2670551

Abstract

A reverse-phase HPLC System for isolation of the water insoluble alpha- and beta-polypeptides of the light-harvesting complex II (LH II) of Rhodopseudomonas (Rps.) palustris without employment of any detergent was developed. The material obtained was of high purity and suitable for direct microsequence analysis. Chromatographic analysis could resolve at least two major beta-polypeptides, beta a and beta b, two major alpha-polypeptides, alpha a and alpha b, and two additional minor polypeptides. N-terminal amino acid sequencing shows that the resolved peaks correspond to different polypeptide species and that the minor species have an N-terminal sequence identical to that of the alpha b polypeptide. An oligonucleotide derived from the amino terminal sequence of the alpha a polypeptide was utilized to screen a genomic library from Rps.palustris. Several independent clones have been characterized by Southern blot and nucleotide sequence analysis. We show that Rps.palustris contains at least four different clusters of beta and alpha genes. Two clones contain sequences potentially coding for beta a-alpha a and beta b-alpha b polypeptides; and two additional clones potentially coding for beta and alpha peptides which we named beta c-alpha c and beta d-alpha d, which did not correspond to the major purified polypeptides. In addition to the protein chemistry data, the conservation at the amino acid level and the presence of canonical ribosomal binding sites upstream of each of the identified genes strongly suggest that all four coding regions are expressed.

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Selected References

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  1. Bensi G., Raugei G., Klefenz H., Cortese R. Structure and expression of the human haptoglobin locus. EMBO J. 1985 Jan;4(1):119–126. doi: 10.1002/j.1460-2075.1985.tb02325.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chory J., Donohue T. J., Varga A. R., Staehelin L. A., Kaplan S. Induction of the photosynthetic membranes of Rhodopseudomonas sphaeroides: biochemical and morphological studies. J Bacteriol. 1984 Aug;159(2):540–554. doi: 10.1128/jb.159.2.540-554.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Drews G. Structure and functional organization of light-harvesting complexes and photochemical reaction centers in membranes of phototrophic bacteria. Microbiol Rev. 1985 Mar;49(1):59–70. doi: 10.1128/mr.49.1.59-70.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dunsmuir P., Smith S. M., Bedbrook J. The major chlorophyll a/b binding protein of petunia is composed of several polypeptides encoded by a number of distinct nuclear genes. J Mol Appl Genet. 1983;2(3):285–300. [PubMed] [Google Scholar]
  5. Firsow N. N., Drews G. Differentiation of the intracytoplasmic membrane of Rhodopseudomonas palustris induced by variations of oxygen partial pressure or light intensity. Arch Microbiol. 1977 Dec 15;115(3):299–306. doi: 10.1007/BF00446456. [DOI] [PubMed] [Google Scholar]
  6. Golden S. S., Brusslan J., Haselkorn R. Expression of a family of psbA genes encoding a photosystem II polypeptide in the cyanobacterium Anacystis nidulans R2. EMBO J. 1986 Nov;5(11):2789–2798. doi: 10.1002/j.1460-2075.1986.tb04569.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hayashi H., Miyao M., Morita S. Absorption and fluorescence spectra of light-harvesting bacteriochlorophyll-protein complexes from Rhodopseudomonas palustris in the near-infrared region. J Biochem. 1982 Mar;91(3):1017–1027. doi: 10.1093/oxfordjournals.jbchem.a133751. [DOI] [PubMed] [Google Scholar]
  8. Hayashi H., Morita S. Near-infrared absorption spectra of light harvesting bacteriochlorophyll protein complexes from Chromatium vinosum. J Biochem. 1980 Nov;88(5):1251–1258. doi: 10.1093/oxfordjournals.jbchem.a133093. [DOI] [PubMed] [Google Scholar]
  9. Hayashi H., Nakano M., Morita S. Comparative studies of protein properties and bacteriochlorophyll contents of bacteriochlorophyll-protein complexes from spectrally different types of Rhodopseudomonas palustris. J Biochem. 1982 Dec;92(6):1805–1811. doi: 10.1093/oxfordjournals.jbchem.a134110. [DOI] [PubMed] [Google Scholar]
  10. Kiley P. J., Kaplan S. Cloning, DNA sequence, and expression of the Rhodobacter sphaeroides light-harvesting B800-850-alpha and B800-850-beta genes. J Bacteriol. 1987 Jul;169(7):3268–3275. doi: 10.1128/jb.169.7.3268-3275.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Klug G., Drews G. Construction of a gene bank of Rhodopseudomonas capsulata using a broad host range DNA cloning system. Arch Microbiol. 1984 Nov;139(4):319–325. doi: 10.1007/BF00408373. [DOI] [PubMed] [Google Scholar]
  12. Kohorn B. D., Harel E., Chitnis P. R., Thornber J. P., Tobin E. M. Functional and mutational analysis of the light-harvesting chlorophyll a/b protein of thylakoid membranes. J Cell Biol. 1986 Mar;102(3):972–981. doi: 10.1083/jcb.102.3.972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Mulligan B., Schultes N., Chen L., Bogorad L. Nucleotide sequence of a multiple-copy gene for the B protein of photosystem II of a cyanobacterium. Proc Natl Acad Sci U S A. 1984 May;81(9):2693–2697. doi: 10.1073/pnas.81.9.2693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Paonessa G., Gounari F., Frank R., Cortese R. Purification of a NF1-like DNA-binding protein from rat liver and cloning of the corresponding cDNA. EMBO J. 1988 Oct;7(10):3115–3123. doi: 10.1002/j.1460-2075.1988.tb03178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Schmidt R., Ackermann R., Kratky Z., Wasserman B., Jacobson B. Fast and efficient purification of yeast plasma membranes using cationic silica microbeads. Biochim Biophys Acta. 1983 Jul 27;732(2):421–427. doi: 10.1016/0005-2736(83)90059-7. [DOI] [PubMed] [Google Scholar]
  18. Shiozawa J. A., Cuendet P. A., Drews G., Zuber H. Isolation and characterization of the polypeptide components from light-harvesting pigment-protein complex B800--850 of Rhodopseudomonas capsulata. Eur J Biochem. 1980 Oct;111(2):455–460. doi: 10.1111/j.1432-1033.1980.tb04960.x. [DOI] [PubMed] [Google Scholar]
  19. Tadros M. H., Zuber H., Drews G. The polypeptide components from light-harvesting pigment-protein complex II (B800-850) of Rhodopseudomonas capsulata. Solubilization, purification and sequence studies. Eur J Biochem. 1982 Oct;127(2):315–318. doi: 10.1111/j.1432-1033.1982.tb06872.x. [DOI] [PubMed] [Google Scholar]
  20. Tauschel H. D., Drews G. Thylakoidmorphogenese bei Rhodopseudomonas palustirs. Arch Mikrobiol. 1967;59(4):381–404. [PubMed] [Google Scholar]
  21. Theiler R., Suter F., Wiemken V., Zuber H. The light-harvesting polypeptides of Rhodopseudomonas sphaeroides R-26.1. I. Isolation, purification and sequence analyses. Hoppe Seylers Z Physiol Chem. 1984 Jul;365(7):703–719. doi: 10.1515/bchm2.1984.365.2.703. [DOI] [PubMed] [Google Scholar]
  22. Varga A. R., Staehelin L. A. Pigment-protein complexes from Rhodopseudomonas palustris: isolation, characterization, and reconstitution into liposomes. J Bacteriol. 1985 Mar;161(3):921–927. doi: 10.1128/jb.161.3.921-927.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Whittenbury R., McLee A. G. Rhodopseudomonas palustris and Rh. viridis--photosynthetic budding bacteria. Arch Mikrobiol. 1967;59(1):324–334. doi: 10.1007/BF00406346. [DOI] [PubMed] [Google Scholar]
  24. Youvan D. C., Alberti M., Begusch H., Bylina E. J., Hearst J. E. Reaction center and light-harvesting I genes from Rhodopseudomonas capsulata. Proc Natl Acad Sci U S A. 1984 Jan;81(1):189–192. doi: 10.1073/pnas.81.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Youvan D. C., Bylina E. J., Alberti M., Begusch H., Hearst J. E. Nucleotide and deduced polypeptide sequences of the photosynthetic reaction-center, B870 antenna, and flanking polypeptides from R. capsulata. Cell. 1984 Jul;37(3):949–957. doi: 10.1016/0092-8674(84)90429-x. [DOI] [PubMed] [Google Scholar]
  26. 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]

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