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. 1994 May 1;13(9):2227–2235. doi: 10.1002/j.1460-2075.1994.tb06500.x

ADP-dependent phosphorylation regulates RNA-binding in vitro: implications in light-modulated translation.

A Danon 1, S P Mayfield 1
PMCID: PMC395078  PMID: 8187775

Abstract

Light-regulated translation of chloroplastic mRNAs in the green alga Chlamydomonas reinhardtii requires nuclear encoded factors that interact with the 5'-untranslated region (5'-UTR) of specific mRNAs to enhance their translation. We have previously identified and characterized a set of proteins that bind specifically to the 5'-UTR of the chloroplastic psbA mRNA. Accumulation of these proteins is similar in dark- and light-grown cells, whereas their binding activity is enhanced during growth in the light. We have identified a serine/threonine protein phosphotransferase, associated with the psbA mRNA-binding complex, that utilizes the beta-phosphate of ADP to phosphorylate and inactivate psbA mRNA-binding in vitro. The inactivation of mRNA-binding in vitro is initiated at high ADP levels, levels that are attained in vivo only in dark-grown chloroplasts. These data suggest that the translation of psbA mRNA is attenuated by phosphorylation of the mRNA-binding protein complex in response to a rise in the stromal concentration of ADP upon transfer of cells to dark.

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  1. 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]
  2. Boyle W. J., van der Geer P., Hunter T. Phosphopeptide mapping and phosphoamino acid analysis by two-dimensional separation on thin-layer cellulose plates. Methods Enzymol. 1991;201:110–149. doi: 10.1016/0076-6879(91)01013-r. [DOI] [PubMed] [Google Scholar]
  3. Buchanan B. B. Regulation of CO2 assimilation in oxygenic photosynthesis: the ferredoxin/thioredoxin system. Perspective on its discovery, present status, and future development. Arch Biochem Biophys. 1991 Jul;288(1):1–9. doi: 10.1016/0003-9861(91)90157-e. [DOI] [PubMed] [Google Scholar]
  4. Budde R. J., Ernst S. M., Chollet R. Substrate specificity and regulation of the maize (Zea mays) leaf ADP: protein phosphotransferase catalysing phosphorylation/inactivation of pyruvate, orthophosphate dikinase. Biochem J. 1986 Jun 1;236(2):579–584. doi: 10.1042/bj2360579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burnell J. N., Hatch M. D. Dark-light regulation of pyruvate, Pi dikinase in C4 plants: evidence that the same protein catalyses activation and inactivation. Biochem Biophys Res Commun. 1983 Feb 28;111(1):288–293. doi: 10.1016/s0006-291x(83)80149-1. [DOI] [PubMed] [Google Scholar]
  6. Coughlan S. J., Hind G. Protein kinases of the thylakoid membrane. J Biol Chem. 1986 Oct 25;261(30):14062–14068. [PubMed] [Google Scholar]
  7. Danon A., Mayfield S. P. Light regulated translational activators: identification of chloroplast gene specific mRNA binding proteins. EMBO J. 1991 Dec;10(13):3993–4001. doi: 10.1002/j.1460-2075.1991.tb04974.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Deng X. W., Gruissem W. Constitutive transcription and regulation of gene expression in non-photosynthetic plastids of higher plants. EMBO J. 1988 Nov;7(11):3301–3308. doi: 10.1002/j.1460-2075.1988.tb03200.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Feldhau P., Fröhlich T., Goody R. S., Isakov M., Schirmer R. H. Synthetic inhibitors of adenylate kinases in the assays for ATPases and phosphokinases. Eur J Biochem. 1975 Sep 1;57(1):197–204. doi: 10.1111/j.1432-1033.1975.tb02291.x. [DOI] [PubMed] [Google Scholar]
  10. Fromm H., Devic M., Fluhr R., Edelman M. Control of psbA gene expression: in mature Spirodela chloroplasts light regulation of 32-kd protein synthesis is independent of transcript level. EMBO J. 1985 Feb;4(2):291–295. doi: 10.1002/j.1460-2075.1985.tb03628.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Girard-Bascou J., Pierre Y., Drapier D. A nuclear mutation affects the synthesis of the chloroplast psbA gene production Chlamydomonas reinhardtii. Curr Genet. 1992 Jul;22(1):47–52. doi: 10.1007/BF00351741. [DOI] [PubMed] [Google Scholar]
  12. Gratecos D., Fischer E. H. Adenosine 5'-O(3-thiotriphosphate) in the control of phosphorylase activity. Biochem Biophys Res Commun. 1974 Jun 18;58(4):960–967. doi: 10.1016/s0006-291x(74)80237-8. [DOI] [PubMed] [Google Scholar]
  13. Guitton C., Dorne A. M., Mache R. In organello and in vitro phosphorylation of chloroplast ribosomal proteins. Biochem Biophys Res Commun. 1984 May 31;121(1):297–303. doi: 10.1016/0006-291x(84)90722-8. [DOI] [PubMed] [Google Scholar]
  14. Guitton C., Mache R. Phosphorylation in vitro of the large subunit of the ribulose-1,5-bisphosphate carboxylase and of the glyceraldehyde-3-phosphate dehydrogenase. Eur J Biochem. 1987 Jul 1;166(1):249–254. doi: 10.1111/j.1432-1033.1987.tb13509.x. [DOI] [PubMed] [Google Scholar]
  15. Hampp R., Goller M., Ziegler H. Adenylate Levels, Energy Charge, and Phosphorylation Potential during Dark-Light and Light-Dark Transition in Chloroplasts, Mitochondria, and Cytosol of Mesophyll Protoplasts from Avena sativa L. Plant Physiol. 1982 Feb;69(2):448–455. doi: 10.1104/pp.69.2.448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Heber U. W., Santarius K. A. Compartmentation and reduction of pyridine nucleotides in relation to photosynthesis. Biochim Biophys Acta. 1965 Nov 29;109(2):390–408. doi: 10.1016/0926-6585(65)90166-4. [DOI] [PubMed] [Google Scholar]
  17. Jensen K. H., Herrin D. L., Plumley F. G., Schmidt G. W. Biogenesis of photosystem II complexes: transcriptional, translational, and posttranslational regulation. J Cell Biol. 1986 Oct;103(4):1315–1325. doi: 10.1083/jcb.103.4.1315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Klein R. R., Mason H. S., Mullet J. E. Light-regulated translation of chloroplast proteins. I. Transcripts of psaA-psaB, psbA, and rbcL are associated with polysomes in dark-grown and illuminated barley seedlings. J Cell Biol. 1988 Feb;106(2):289–301. doi: 10.1083/jcb.106.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kuchka M. R., Mayfield S. P., Rochaix J. D. Nuclear mutations specifically affect the synthesis and/or degradation of the chloroplast-encoded D2 polypeptide of photosystem II in Chlamydomonas reinhardtii. EMBO J. 1988 Feb;7(2):319–324. doi: 10.1002/j.1460-2075.1988.tb02815.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Lin Z. F., Lucero H. A., Racker E. Protein kinases from spinach chloroplasts. I. Purification and identification of two distinct protein kinases. J Biol Chem. 1982 Oct 25;257(20):12153–12156. [PubMed] [Google Scholar]
  22. Malnoë P., Mayfield S. P., Rochaix J. D. Comparative analysis of the biogenesis of photosystem II in the wild-type and Y-1 mutant of Chlamydomonas reinhardtii. J Cell Biol. 1988 Mar;106(3):609–616. doi: 10.1083/jcb.106.3.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Michel H., Hunt D. F., Shabanowitz J., Bennett J. Tandem mass spectrometry reveals that three photosystem II proteins of spinach chloroplasts contain N-acetyl-O-phosphothreonine at their NH2 termini. J Biol Chem. 1988 Jan 25;263(3):1123–1130. [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. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  27. Palczewski K., Kahn N., Hargrave P. A. Nucleoside inhibitors of rhodopsin kinase. Biochemistry. 1990 Jul 3;29(26):6276–6282. doi: 10.1021/bi00478a024. [DOI] [PubMed] [Google Scholar]
  28. Rochaix J. D., Kuchka M., Mayfield S., Schirmer-Rahire M., Girard-Bascou J., Bennoun P. Nuclear and chloroplast mutations affect the synthesis or stability of the chloroplast psbC gene product in Chlamydomonas reinhardtii. EMBO J. 1989 Apr;8(4):1013–1021. doi: 10.1002/j.1460-2075.1989.tb03468.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stitt M., Wirtz W., Heldt H. W. Metabolite levels during induction in the chloroplast and extrachloroplast compartments of spinach protoplasts. Biochim Biophys Acta. 1980 Nov 5;593(1):85–102. doi: 10.1016/0005-2728(80)90010-9. [DOI] [PubMed] [Google Scholar]
  30. Sun G., Bailey D., Jones M. W., Markwell J. Chloroplast thylakoid protein phosphatase is a membrane surface-associated activity. Plant Physiol. 1989 Jan;89(1):238–243. doi: 10.1104/pp.89.1.238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yu X., Egelman E. H. Direct visualization of dynamics and co-operative conformational changes within RecA filaments that appear to be associated with the hydrolysis of adenosine 5'-O-(3-thiotriphosphate). J Mol Biol. 1992 May 5;225(1):193–216. doi: 10.1016/0022-2836(92)91036-o. [DOI] [PubMed] [Google Scholar]

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