Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1985 Jun;5(6):1370–1378. doi: 10.1128/mcb.5.6.1370

Structure and developmental regulation of a wheat gene encoding the major chlorophyll a/b-binding polypeptide.

G K Lamppa, G Morelli, N H Chua
PMCID: PMC366866  PMID: 3897835

Abstract

A genomic clone for a major chlorophyll a/b-binding polypeptide of the light-harvesting complex has been sequenced from wheat. This gene, whAB1.6, encodes a 70-nucleotide 5'-nontranslated spacer, a 34-amino-acid NH2-terminal extension, i.e., the transit peptide, and a mature coding protein of 232 amino acid residues. The exact molecular weight of the precursor polypeptide is 28,560. The transit peptide is basic and is rich in serines. No intervening sequences are found in this gene. The transcription start site of the whAB1.6 gene occurs at AAAC as determined by S1 nuclease analysis. Putative regulatory sequences occur upstream of the gene at -25 (TTTAAATA) and at -72 (CCAACCA). Northern blots show a single RNA species estimated to be 1,100 nucleotides. Heterogeneity of the RNA population is demonstrated in S1 nuclease analyses with a 5'-end-labeled fragment that extends 191 nucleotides into the mature protein coding sequence. At least seven different transcripts can be recognized. The highest levels of RNA transcribed from the whAB1.6 gene are found in the basal segments of the wheat leaf, whereas other chlorophyll a/b-binding transcripts in the cell show a different pattern of abundance. As a control, we show that roots do not contain chlorophyll a/b-binding RNA. The most abundant RNA species shows an interrupted homology with the whAB1.6 gene at the start of the mature protein coding sequence; another species shows homology beginning at the start of the transit peptide and does not include the nontranslated region. Chlorophyll a/b-binding polypeptides accumulate toward the tip of the leaf as shown by Western blot analysis of total thylakoid proteins.

Full text

PDF
1370

Images in this article

1373Image
on p.1373
1374Image
on p.1374
1374Image
on p.1374
1375Image
on p.1375
1376Image
on p.1376

Selected References

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

  1. Apel K. Phytochrome-induced appearance of mRNA activity for the apoprotein of the light-harvesting chlorophyll a/b protein of barley (Hordeum vulgare). Eur J Biochem. 1979 Jun;97(1):183–188. doi: 10.1111/j.1432-1033.1979.tb13101.x. [DOI] [PubMed] [Google Scholar]
  2. Bennett J. Regulation of photosynthesis by reversible phosphorylation of the light-harvesting chlorophyll a/b protein. Biochem J. 1983 Apr 15;212(1):1–13. doi: 10.1042/bj2120001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett J., Steinback K. E., Arntzen C. J. Chloroplast phosphoproteins: regulation of excitation energy transfer by phosphorylation of thylakoid membrane polypeptides. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5253–5257. doi: 10.1073/pnas.77.9.5253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  6. Boffey S. A., Ellis J. R., Selldén G., Leech R. M. Chloroplast Division and DNA Synthesis in Light-grown Wheat Leaves. Plant Physiol. 1979 Sep;64(3):502–505. doi: 10.1104/pp.64.3.502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  8. Carmichael G. G., McMaster G. K. The analysis of nucleic acids in gels using glyoxal and acridine orange. Methods Enzymol. 1980;65(1):380–391. doi: 10.1016/s0076-6879(80)65049-6. [DOI] [PubMed] [Google Scholar]
  9. Cashmore A. R. Structure and expression of a pea nuclear gene encoding a chlorophyll a/b-binding polypeptide. Proc Natl Acad Sci U S A. 1984 May;81(10):2960–2964. doi: 10.1073/pnas.81.10.2960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  11. Coruzzi G., Broglie R., Cashmore A., Chua N. H. Nucleotide sequences of two pea cDNA clones encoding the small subunit of ribulose 1,5-bisphosphate carboxylase and the major chlorophyll a/b-binding thylakoid polypeptide. J Biol Chem. 1983 Feb 10;258(3):1399–1402. [PubMed] [Google Scholar]
  12. Coruzzi G., Broglie R., Edwards C., Chua N. H. Tissue-specific and light-regulated expression of a pea nuclear gene encoding the small subunit of ribulose-1,5-bisphosphate carboxylase. EMBO J. 1984 Aug;3(8):1671–1679. doi: 10.1002/j.1460-2075.1984.tb02031.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cuming A. C., Bennett J. Biosynthesis of the light-harvesting chlorophyll a/b protein. Control of messenger RNA activity by light. Eur J Biochem. 1981 Aug;118(1):71–80. doi: 10.1111/j.1432-1033.1981.tb05487.x. [DOI] [PubMed] [Google Scholar]
  14. Dean C., Leech R. M. Genome Expression during Normal Leaf Development : I. CELLULAR AND CHLOROPLAST NUMBERS AND DNA, RNA, AND PROTEIN LEVELS IN TISSUES OF DIFFERENT AGES WITHIN A SEVEN-DAY-OLD WHEAT LEAF. Plant Physiol. 1982 Apr;69(4):904–910. doi: 10.1104/pp.69.4.904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Delepelaire P., Chua N. H. Electrophoretic purification of chlorophyll a/b-protein complexes from Chlamydomonas reinhardtii and spinach and analysis of their polypeptide compositions. J Biol Chem. 1981 Sep 10;256(17):9300–9307. [PubMed] [Google Scholar]
  16. 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]
  17. Gallagher T. F., Ellis R. J. Light-stimulated transcription of genes for two chloroplast polypeptides in isolated pea leaf nuclei. EMBO J. 1982;1(12):1493–1498. doi: 10.1002/j.1460-2075.1982.tb01345.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gerken S. C., Arfin S. M. Threonyl-tRNA synthetase from Chinese hamster ovary cells is phosphorylated on serine. J Biol Chem. 1984 Sep 25;259(18):11160–11161. [PubMed] [Google Scholar]
  19. Glazer A. N. Comparative biochemistry of photosynthetic light-harvesting systems. Annu Rev Biochem. 1983;52:125–157. doi: 10.1146/annurev.bi.52.070183.001013. [DOI] [PubMed] [Google Scholar]
  20. Lindberg U., Persson T. Isolation of mRNA from KB-cells by affinity chromatography on polyuridylic acid covalently linked to Sepharose. Eur J Biochem. 1972 Dec 4;31(2):246–254. doi: 10.1111/j.1432-1033.1972.tb02527.x. [DOI] [PubMed] [Google Scholar]
  21. Makino A., Mae T., Ohira K. Photosynthesis and Ribulose 1,5-Bisphosphate Carboxylase in Rice Leaves: Changes in Photosynthesis and Enzymes Involved in Carbon Assimilation from Leaf Development through Senescence. Plant Physiol. 1983 Dec;73(4):1002–1007. doi: 10.1104/pp.73.4.1002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McKeown M., Firtel R. A. Differential expression and 5' end mapping of actin genes in Dictyostelium. Cell. 1981 Jun;24(3):799–807. doi: 10.1016/0092-8674(81)90105-7. [DOI] [PubMed] [Google Scholar]
  24. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  25. Mullet J. E. The amino acid sequence of the polypeptide segment which regulates membrane adhesion (grana stacking) in chloroplasts. J Biol Chem. 1983 Aug 25;258(16):9941–9948. [PubMed] [Google Scholar]
  26. Nevins J. R. The pathway of eukaryotic mRNA formation. Annu Rev Biochem. 1983;52:441–466. doi: 10.1146/annurev.bi.52.070183.002301. [DOI] [PubMed] [Google Scholar]
  27. Rimpau J., Smith D., Flavell R. Sequence organisation analysis of the wheat and rye genomes by interspecies DNA/DNA hybridisation. J Mol Biol. 1978 Aug 15;123(3):327–359. doi: 10.1016/0022-2836(78)90083-9. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Scheidtmann K. H., Echle B., Walter G. Simian virus 40 large T antigen is phosphorylated at multiple sites clustered in two separate regions. J Virol. 1982 Oct;44(1):116–133. doi: 10.1128/jvi.44.1.116-133.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schmidt G. W., Bartlett S. G., Grossman A. R., Cashmore A. R., Chua N. H. Biosynthetic pathways of two polypeptide subunits of the light-harvesting chlorophyll a/b protein complex. J Cell Biol. 1981 Nov;91(2 Pt 1):468–478. doi: 10.1083/jcb.91.2.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  32. Staehelin L. A., Arntzen C. J. Regulation of chloroplast membrane function: protein phosphorylation changes the spatial organization of membrane components. J Cell Biol. 1983 Nov;97(5 Pt 1):1327–1337. doi: 10.1083/jcb.97.5.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stiekema W. J., Wimpee C. F., Silverthorne J., Tobin E. M. Phytochrome Control of the Expression of Two Nuclear Genes Encoding Chloroplast Proteins in Lemna gibba L. G-3. Plant Physiol. 1983 Jul;72(3):717–724. doi: 10.1104/pp.72.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tsunewaki K., Ogihara Y. The Molecular Basis of Genetic Diversity among Cytoplasms of TRITICUM and AEGILOPS Species. II. on the Origin of Polyploid Wheat Cytoplasms as Suggested by Chloroplast DNA Restriction Fragment Patterns. Genetics. 1983 May;104(1):155–171. doi: 10.1093/genetics/104.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES