Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Dec;79(24):7699–7703. doi: 10.1073/pnas.79.24.7699

Nucleotide sequence of the gene for the Mr 32,000 thylakoid membrane protein from Spinacia oleracea and Nicotiana debneyi predicts a totally conserved primary translation product of Mr 38,950

Gerard Zurawski *,†,, Hans J Bohnert *,†,§, Paul R Whitfeld *,, Warwick Bottomley *,
PMCID: PMC347415  PMID: 16593262

Abstract

The gene for the so-called Mr 32,000 rapidly labeled photosystem II thylakoid membrane protein (here designated psbA) of spinach (Spinacia oleracea) chloroplasts is located on the chloroplast DNA in the large single-copy region immediately adjacent to one of the inverted repeat sequences. In this paper we show that the size of the mRNA for this protein is ≈ 1.25 kilobases and that the direction of transcription is towards the inverted repeat unit. The nucleotide sequence of the gene and its flanking regions is presented. The only large open reading frame in the sequence codes for a protein of Mr 38,950. The nucleotide sequence of psbA from Nicotiana debneyi also has been determined, and comparison of the sequences from the two species shows them to be highly conserved (>95% homology) throughout the entire reading frame. Conservation of the amino acid sequence is absolute, there being no changes in a total of 353 residues. This leads us to conclude that the primary translation product of psbA must be a protein of Mr 38,950. The protein is characterized by the complete absence of lysine residues and is relatively rich in hydrophobic amino acids, which tend to be clustered. Transcription of spinach psbA starts about 86 base pairs before the first ATG codon. Immediately upstream from this point there is a sequence typical of that found in E. coli promoters. An almost identical sequence occurs in the equivalent region of N. debneyi DNA.

Keywords: chloroplast DNA, psbA mRNA, amino acid sequence

Full text

PDF
7701

Images in this article

Selected References

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

  1. 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]
  2. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  3. Borer P. N., Dengler B., Tinoco I., Jr, Uhlenbeck O. C. Stability of ribonucleic acid double-stranded helices. J Mol Biol. 1974 Jul 15;86(4):843–853. doi: 10.1016/0022-2836(74)90357-x. [DOI] [PubMed] [Google Scholar]
  4. Bottomley W., Spencer D., Whitfeld P. R. Protein synthesis in isolated spinach chloroplasts: comparison of light-driven and ATP-driven synthesis. Arch Biochem Biophys. 1974 Sep;164(1):106–117. doi: 10.1016/0003-9861(74)90012-5. [DOI] [PubMed] [Google Scholar]
  5. Darley-Usmar V. M., Fuller S. D. Mr-values of mature subunits I and III of beef heart cytochrome c oxidase in relationship to nucleotide sequences of their genes. FEBS Lett. 1981 Nov 30;135(1):164–166. doi: 10.1016/0014-5793(81)80968-4. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Grebanier A. E., Coen D. M., Rich A., Bogorad L. Membrane proteins synthesized but not processed by isolated maize chloroplasts. J Cell Biol. 1978 Sep;78(3):734–746. doi: 10.1083/jcb.78.3.734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hartley M. R., Wheeler A., Ellis R. J. Protein synthesis in chloroplasts. V. Translation of messenger RNA for the large subunit of fraction I protein in a heterologous cell-free system. J Mol Biol. 1975 Jan 5;91(1):67–77. doi: 10.1016/0022-2836(75)90372-1. [DOI] [PubMed] [Google Scholar]
  9. Hartley S. E., McBeath S. Cytogenetic follow-up in chronic myeloid leukemia. Cancer Genet Cytogenet. 1981 Jan;3(1):37–46. doi: 10.1016/0165-4608(81)90054-6. [DOI] [PubMed] [Google Scholar]
  10. Herrmann R., Neugebauer K., Pirkl E., Zentgraf H., Schaller H. Conversion of bacteriophage fd into an efficient single-stranded DNA vector system. Mol Gen Genet. 1980 Jan;177(2):231–242. doi: 10.1007/BF00267434. [DOI] [PubMed] [Google Scholar]
  11. Hoffman-Falk H., Mattoo A. K., Marder J. B., Edelman M., Ellis R. J. General occurrence and structural similarity of the rapidly synthesized, 32,000-dalton protein of the chloroplast membrane. J Biol Chem. 1982 Apr 25;257(8):4583–4587. [PubMed] [Google Scholar]
  12. Kuwabara T., Murata N. Purification and characterization of 33 kilodalton protein of spinach chloroplasts. Biochim Biophys Acta. 1979 Dec 14;581(2):228–236. doi: 10.1016/0005-2795(79)90242-3. [DOI] [PubMed] [Google Scholar]
  13. Link G. Cloning and mapping of the chloroplast DNA sequences for two messenger RNAs from mustard (Sinapis alba L.). Nucleic Acids Res. 1981 Aug 11;9(15):3681–3694. doi: 10.1093/nar/9.15.3681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mattoo A. K., Pick U., Hoffman-Falk H., Edelman M. The rapidly metabolized 32,000-dalton polypeptide of the chloroplast is the "proteinaceous shield" regulating photosystem II electron transport and mediating diuron herbicide sensitivity. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1572–1576. doi: 10.1073/pnas.78.3.1572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Morgenthaler J. J., Mendiola-Morgenthaler L. Synthesis of soluble, thylakoid, and envelope membrane proteins by spinach chloroplasts purified from gradients. Arch Biochem Biophys. 1976 Jan;172(1):51–58. doi: 10.1016/0003-9861(76)90046-1. [DOI] [PubMed] [Google Scholar]
  17. Palmer J. D. Physical and gene mapping of chloroplast DNA from Atriplex triangularis and Cucumis sativa. Nucleic Acids Res. 1982 Mar 11;10(5):1593–1605. doi: 10.1093/nar/10.5.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pfister K., Steinback K. E., Gardner G., Arntzen C. J. Photoaffinity labeling of an herbicide receptor protein in chloroplast membranes. Proc Natl Acad Sci U S A. 1981 Feb;78(2):981–985. doi: 10.1073/pnas.78.2.981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Schreier P. H., Cortese R. A fast and simple method for sequencing DNA cloned in the single-stranded bacteriophage M13. J Mol Biol. 1979 Mar 25;129(1):169–172. doi: 10.1016/0022-2836(79)90068-8. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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 Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES