Photosynthesis Research

, Volume 49, Issue 3, pp 223–235 | Cite as

The purple photosynthetic bacterium Rhodopseudomonas acidophila contains multiple puc peripheral antenna complex (LH2) genes: Cloning and initial characterisation of four β/α pairs

  • Alastair T. Gardiner
  • R. Christopher MacKenzie
  • Stuart J. Barrett
  • Kim Kaiser
  • Richard J. Cogdell
Regular Paper

Abstract

The genome of the purple non-sulphur photosynthetic bacterium Rhodopseudomonas acidophila has been found to contain multiple copies of puc light-harvesting (LH2) peripheral antenna complex genes. Three wild-type isolates each exhibiting dissimilar peripheral antenna complex phenotypes in response to growth at reduced light intensity, were found to contain different numbers of these genes. Twenty-three puc cross-hybridising clones were isolated from a genomic library constructed from Rhodopseudomonas acidophila strain 7050; two of which were examined further; 2.6kb from one clone was sequenced and found to contain three β/α gene pairs designated puc1BA, puc2BA and puc3BA. The putative translated polypeptides are very like, but not identical to those from B800–820 complexes and upstream sequence homologies suggests that this treble gene cluster has arisen through a relatively recent gene duplication event. From the other clone 0.6kb was sequenced and found to contain a further gene pair, puc4BA, which is capable of encoding apoproteins for a B800-850-like complex. When the cells are grown at ‘high’ or ‘low’ light intensity Northern analyses showed that only puc4BA is expressed under ‘high’ light conditions. Furthermore, pucBA mRNA transcripts were detected in all three species in the range 500–780 nt. In Rhodopseudomonas acidophila post-transcriptional regulatory mechanisms also play a role in determining the amount of peripheral antenna present in the intra-cytoplasmic membrane.

Key words

light-harvesting multigene family 

Abbreviations

Bchl-

bacteriochlorophyll

ICM-

intra-cytoplasmic membrane

LH1-

core light-harvesting antenna complex

LH2-

peripheral light-harvesting antenna complex

NIR-

near infra-red

ORF-

open reading frame

PSU-

photosynthetic unit

RC-

reaction centre

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Aargard J and Sistrom WR (1972) Control of synthesis of reaction centre bacteriochlorophyll in photosynthetic bacteria. Photochem Photobiol 15: 209–225Google Scholar
  2. Ashby MK, Coomber SA and Hunter CN (1987) Cloning, nucleotide sequence and transfer of genes for the B800–850 light harvesting complex of Rhodobacter sphaeroides. FEBS Lett 213: 245–248Google Scholar
  3. Chomczynski P and Sacchi N (1987) Single step method of RNA isolation by acid Guanidium Thiocyanate-Phenol-Chloroform extraction. Anal Biochem 162: 156–159Google Scholar
  4. Cogdell RJ, Durant I, Valentine J, Lindsay JG and Schmidt K (1983) The isolation and partial characterisation of the light-harvesting pigment protein complement from Rhodopseudomonas acidophila. Biochim Biophys Acta 722: 427–455Google Scholar
  5. Deinum G, Otte SCM, Gardiner AT, Aartsma TJ, Cogdell RJ and Amesz J (1991) Antenna organisation of Rhodopseudomonas acidophila: A study of the excitation migration. Biochim Biophys Acta 1060: 125–131Google Scholar
  6. Fowler GJS, Gardiner AT, MacKenzie RC, Barrett SJ, Simmons AE, Westerhuis WHJ, Cogdell RJ and Hunter CN (1995) Heterologous expression of genes encoding bacterial light-harvesting complexes in Rhodobacter sphaeroides. J Biol Chem 270: 1–8Google Scholar
  7. Fowler GJS, Sockalingum GD, Robert B and Hunter CN (1994) Blue shifts in bacteriochorophyll absorbance correlate with changed hydrogen bonding patterns in light-harvesting 2 mutants of Rhodobacter sphaeroides with alterations at α-Tyr-44 and α-Tyr-45. Biochem J 299: 695–700Google Scholar
  8. Fowler GJS, Visschers RW, Grief GG, van Grondelle R and Hunter CN (1992) Genetically modified photosynthetic antenna complexes with blueshifted absorbance bands. Nature 355: 848–850Google Scholar
  9. Gardiner AT, Cogdell RJ and Takaichi S (1993) The effect of growth conditions on the light-harvesting apparatus in Rhodopseudomonas acidophila. Photosynth Res 38: 159–167Google Scholar
  10. Guthrie N, MacDermott G, Cogdell RJ, Freer AA, Isaacs NW, Hawthornthwaite AM, Halloren E, Lindsay JG, (1992) Crystallisation of the B800–820 light-harvesting complex from Rhodopseudomonas acidophila strain 7050. J Mol Biol 224: 527–528Google Scholar
  11. Heinemeyer EA and Schmidt K (1983) Changes in carotenoid biosynthesis caused by variations of growth conditions in cultures of Rhodopseudomonas acidophila strain 7050. Arch Microbiol 134: 217–221Google Scholar
  12. Kaplan S and Donohue TJ (1993) Genetic analysis of photosynthetic membrane biosynthesis in Rhodobacter sphaeroides. In: Deisenhofer J and Norris JR (eds) The Photosynthetic Reaction Centre, Vol 1, pp 101–131. Academic Press, San DiegoGoogle Scholar
  13. Kiley PJ and Kaplan S (1987) Cloning, DNA sequence and expression of the Rhodobacter sphaeroides light-harvesting B800–850(alpha) and B800–850(beta) genes. J Bacteriol 169: 3268–3275Google Scholar
  14. Koepke J, Hu X, Münke C, Schulten K and Michel H (1996) The crystal stucture of the light-harvesting complex II (B800–850) from Rhodospirillum molischianum, Structure 4: 581–597.Google Scholar
  15. Mason PJ and Williams JG (1985) Hybridisation in the analysis of recombinant DNA. In: Hames BD and Higgins SJ (eds) Nucleic Acid Hybridisation: A Practical Approach, pp 113–137. IRL Press, Oxford and Washington, DCGoogle Scholar
  16. McDermott G, Prince SM, Freer AA, Hawthorthwaite-Lawless AM, Papiz MZ, Cogdell RJ and Isaacs NW (1995) Crystal structure of an integral membrane light-harvesting complexc from photosynthetic bacteria. Nature 374: 517–521Google Scholar
  17. Pfennig N (1969) Rhodopseudomonas acidophila sp. n., a new species of the budding purple non-sulphur bacteria. J Bacteriol 99: 597–602Google Scholar
  18. Sambrook J, Fritsch EF and Maniatis T (1989) In: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbour Laboratory Press. Cold Spring Harbour, New YorkGoogle Scholar
  19. Sanger F, Nicklen S and Coulsen AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci 74: 5463–5467Google Scholar
  20. Schumacher A and Drews G (1979) Effects of light intensity on membrane differentiation in Rhodopseudomonas capsulata. Biochim Biophys Acta 501: 417–428Google Scholar
  21. Stadtwald-Demchick R, Turner FR and Gest H (1990) Rhodopseudomonas cryptolactis sp. nov., a new thermotolerant species of budding phototrophic purple bacteria. FEMS Microbiol Lett 71: 117–122Google Scholar
  22. Tadros MH, Katsiou E, Hoon MA, Yurkova N and Ramji DP (1993) Cloning of a new antenna gene cluster and expression analysis of the antenna gene family of Rhodopseudomonas palustris. Eur J Biochem 217: 867–875Google Scholar
  23. Whittaker PA, Campbell AJB, Southern EM and Murray NM (1987) Enhanced recovery and restriction mapping of DNA fragments cloned in a lambda vector. Nucl Acid Res 16: 6725–6736Google Scholar
  24. Willems A, Gillis M and De Ley J (1991) Transfer of Rhodocyclus gelatinosus to Rubrivivax gelatinosus gen. nov. comb. nov., and phylogenetic relationships with Leptothrix, Sphaerotilus natans, Pseudomonas saccharophila and Alcaligenes latus. Int J System Bacteriol 41: 65–73Google Scholar
  25. Youvan DC and Ismail S (1985) Light-harvesting II (B800–850 complex) structural genes from Rhodopseudomonas capsulata. Proc Natl Acad Sci 82: 58–82Google Scholar
  26. Zuber H (1990) Considerations on the structural principles of the antenna complexes of phototrophic bacteria. In: Drews G and Dawes EA (eds) Molecular Biology of Membrane-Bound Complexes in Phototrophic Bacteria, pp 161–180, FEMS Symposium No 53. Plenum Press New York and LondonGoogle Scholar
  27. Zuber H. and Brunisholz R.A. (1991) Structure and function of antenna polypeptides and chlorophyll-protein complexes: principles and variability. In: Scheer H (ed) Chlorophylls, 627–703. CRC Press, Boca RatonGoogle Scholar
  28. Zucconi AP and Beatty JT (1988) Post-trancriptional regulation by light of the steady state levels of mature B800–850 light-harvesting complexes in Rhodobacter capsulatus. J Bacteriol 170: 877–882Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Alastair T. Gardiner
    • 1
  • R. Christopher MacKenzie
    • 1
  • Stuart J. Barrett
    • 1
  • Kim Kaiser
    • 2
  • Richard J. Cogdell
    • 1
  1. 1.Department of Biochemistry, School of Biological SciencesUniversity of GlasgowGlasgowScotland
  2. 2.Department of Genetics, School of Biological SciencesUniversity of GlasgowGlasgowScotland

Personalised recommendations