Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1983;2(12):2229–2237. doi: 10.1002/j.1460-2075.1983.tb01728.x

Localization of the genes for the two chlorophyll a-conjugated polypeptides (mol. wt. 51 and 44 kd) of the photosystem II reaction center on the spinach plastid chromosome

Peter Westhoff 1,*, Juliane Alt 1, Reinhold G Herrmann 1
PMCID: PMC555439  PMID: 16453486

Abstract

A core particle of the water-oxidizing photosystem II reaction center has been prepared from stacked spinach thylakoid membranes by a procedure involving extraction with the non-ionic detergent dodecyl-β-D-maltoside and centrifugation in sucrose gradients. The protein-pigment complex consists of at least four polypeptide species: two components with mol. wts. of 51 and 44 kd which are conjugated with chlorophyll a and β-carotene, the herbicide-binding protein of mol. wt. 32 kd and cytochrome b 559 (11 kd). The genes for the 51-and 44-kd polypeptides have been located on the circular 150-kbp spinach plastid chromosome. They were identified by hybrid-selection mapping, in vitro transcription-translation of recombinant DNAs and specific antisera which were used to characterize the translation products. The plastid chromosome carries one uninterrupted copy for each of these genes in its large single-copy region. The gene for the 51-kd protein (which probably bears the P680 reaction center chlorophyll a) is located in close proximity to the gene for cytochrome b6, and some 70 kbp away from the gene for the `32-kd' herbicide-binding protein of the reducing side of photosystem II. The gene for the 44-kd protein is situated halfway between these two genes adjacent to the gene for the P700 chlorophyll a apoprotein of the photosystem I reaction center. Both photosystem II genes are transcribed into discrete RNA species in the same direction but from the opposite strand as the gene for the `32-kd' protein.

Keywords: photosystem II genes, chlorophyll a-binding polypeptides, thylakoid membrane, plastid DNA, transcripts, spinach

Full text

PDF
2229

Images in this article

Selected References

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

  1. Alt J., Westhoff P., Sears B. B., Nelson N., Hurt E., Hauska G., Herrmann R. G. Genes and transcripts for the polypeptides of the cytochrome b6/f complex from spinach thylakoid membranes. EMBO J. 1983;2(6):979–986. doi: 10.1002/j.1460-2075.1983.tb01531.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Apel K., Kloppstech K. The plastid membranes of barley (Hordeum vulgare). Light-induced appearance of mRNA coding for the apoprotein of the light-harvesting chlorophyll a/b protein. Eur J Biochem. 1978 Apr 17;85(2):581–588. doi: 10.1111/j.1432-1033.1978.tb12273.x. [DOI] [PubMed] [Google Scholar]
  3. Arrand J. R., Myers P. A., Roberts R. J. A new restriction endonuclease from Streptomyces albus G. J Mol Biol. 1978 Jan 5;118(1):127–135. doi: 10.1016/0022-2836(78)90249-8. [DOI] [PubMed] [Google Scholar]
  4. Bedbrook J. R., Link G., Coen D. M., Bogorad L. Maize plastid gene expressed during photoregulated development. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3060–3064. doi: 10.1073/pnas.75.7.3060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bengis C., Nelson N. Purification and properties of the photosystem I reaction center from chloroplasts. J Biol Chem. 1975 Apr 25;250(8):2783–2788. [PubMed] [Google Scholar]
  6. Bünemann H., Westhoff P., Herrmann R. G. Immobilization of denatured DNA to macroporous supports: I. Efficiency of different coupling procedures. Nucleic Acids Res. 1982 Nov 25;10(22):7163–7180. doi: 10.1093/nar/10.22.7163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chua N. H., Bennoun P. Thylakoid membrane polypeptides of Chlamydomonas reinhardtii: wild-type and mutant strains deficient in photosystem II reaction center. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2175–2179. doi: 10.1073/pnas.72.6.2175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chua N. H., Blomberg F. Immunochemical studies of thylakoid membrane polypeptides from spinach and Chlamydomonas reinhardtii. A modified procedure for crossed immunoelectrophoresis of dodecyl sulfate.protein complexes. J Biol Chem. 1979 Jan 10;254(1):215–223. [PubMed] [Google Scholar]
  9. Chua N. H., Gillham N. W. The sites of synthesis of the principal thylakoid membrane polypeptides in Chlamydomonas reinhardtii. J Cell Biol. 1977 Aug;74(2):441–452. doi: 10.1083/jcb.74.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Delepelaire P., Chua N. H. Lithium dodecyl sulfate/polyacrylamide gel electrophoresis of thylakoid membranes at 4 degrees C: Characterizations of two additional chlorophyll a-protein complexes. Proc Natl Acad Sci U S A. 1979 Jan;76(1):111–115. doi: 10.1073/pnas.76.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Driesel A. J., Crouse E. J., Gordon K., Bohnert H. J., Herrmann R. G., Steinmetz A., Mubumbila M., Keller M., Burkard G., Weil J. H. Fractionation and identification of spinach chloroplast transfer RNAs and mapping of their genes on the restriction map of chloroplast DNA. Gene. 1979 Aug;6(4):285–306. doi: 10.1016/0378-1119(79)90070-2. [DOI] [PubMed] [Google Scholar]
  12. Driesel A. J., Speirs J., Bohnert H. J. Spinach chloroplast mRNA for a 32 000 dalton polypeptide: size and localization on the physical map of the chloroplast DNA. Biochim Biophys Acta. 1980 Dec 11;610(2):297–310. doi: 10.1016/0005-2787(80)90011-8. [DOI] [PubMed] [Google Scholar]
  13. Grunstein M., Wallis J. Colony hybridization. Methods Enzymol. 1979;68:379–389. doi: 10.1016/0076-6879(79)68027-8. [DOI] [PubMed] [Google Scholar]
  14. Herrmann R. G., Possingham J. V. Plastid DNA-the plastome. Results Probl Cell Differ. 1980;10:45–96. doi: 10.1007/978-3-540-38255-3_3. [DOI] [PubMed] [Google Scholar]
  15. Herrmann R. G., Whitfeld P. R., Bottomley W. Construction of a SalI/PstI restriction map of spinach chloroplast DNA using low-gelling-temperature-agarose electrophoresis. Gene. 1980 Jan;8(2):179–191. doi: 10.1016/0378-1119(80)90036-0. [DOI] [PubMed] [Google Scholar]
  16. Holmes W. M., Platt T., Rosenberg M. Termination of transcription in E. coli. Cell. 1983 Apr;32(4):1029–1032. doi: 10.1016/0092-8674(83)90287-8. [DOI] [PubMed] [Google Scholar]
  17. Hurt E., Hauska G. A cytochrome f/b6 complex of five polypeptides with plastoquinol-plastocyanin-oxidoreductase activity from spinach chloroplasts. Eur J Biochem. 1981 Jul;117(3):591–595. doi: 10.1111/j.1432-1033.1981.tb06379.x. [DOI] [PubMed] [Google Scholar]
  18. Katz L., Kingsbury D. T., Helinski D. R. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex. J Bacteriol. 1973 May;114(2):577–591. doi: 10.1128/jb.114.2.577-591.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mullet J. E., Burke J. J., Arntzen C. J. Chlorophyll proteins of photosystem I. Plant Physiol. 1980 May;65(5):814–822. doi: 10.1104/pp.65.5.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  21. Pick U., Racker E. Purification and reconstitution of the N,N'-dicyclohexylcarbodiimide-sensitive ATPase complex from spinach chloroplasts. J Biol Chem. 1979 Apr 25;254(8):2793–2799. [PubMed] [Google Scholar]
  22. Renger G. Studies on the structural and functional organization of system II of photosynthesis. The use of trypsin as a structurally selective inhibitor at the outer surface of the thylakoid membrane. Biochim Biophys Acta. 1976 Aug 13;440(2):287–300. doi: 10.1016/0005-2728(76)90063-3. [DOI] [PubMed] [Google Scholar]
  23. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  24. Smith H. O., Birnstiel M. L. A simple method for DNA restriction site mapping. Nucleic Acids Res. 1976 Sep;3(9):2387–2398. doi: 10.1093/nar/3.9.2387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Steinback K. E., McIntosh L., Bogorad L., Arntzen C. J. Identification of the triazine receptor protein as a chloroplast gene product. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7463–7467. doi: 10.1073/pnas.78.12.7463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Whitfeld P. R., Herrmann R. G., Bottomley W. Mapping of the ribosomal RNA genes on spinach chloroplast DNA. Nucleic Acids Res. 1978 Jun;5(6):1741–1751. doi: 10.1093/nar/5.6.1741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zielinski R. E., Price C. A. Synthesis of thylakoid membrane proteins by chloroplasts isolated from spinach. Cytochrome b559 and P700-chlorophyll a-protein. J Cell Biol. 1980 May;85(2):435–445. doi: 10.1083/jcb.85.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zurawski G., Bohnert H. J., Whitfeld P. R., Bottomley W. Nucleotide sequence of the gene for the M(r) 32,000 thylakoid membrane protein from Spinacia oleracea and Nicotiana debneyi predicts a totally conserved primary translation product of M(r) 38,950. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7699–7703. doi: 10.1073/pnas.79.24.7699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zurawski G., Bottomley W., Whitfeld P. R. Structures of the genes for the beta and epsilon subunits of spinach chloroplast ATPase indicate a dicistronic mRNA and an overlapping translation stop/start signal. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6260–6264. doi: 10.1073/pnas.79.20.6260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zurawski G., Perrot B., Bottomley W., Whitfeld P. R. The structure of the gene for the large subunit of ribulose 1,5-bisphosphate carboxylase from spinach chloroplast DNA. Nucleic Acids Res. 1981 Jul 24;9(14):3251–3270. doi: 10.1093/nar/9.14.3251. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES