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Assembly of the barley light-harvesting chlorophyll a/b proteins in barley etiochloroplasts involves processing of the precursor on thylakoids

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Abstract

A barley gene encoding the major light-harvesting chlorophyll a/b-binding protein (LHCP) has been sequenced and then expressed in vitro to produce a labelled LHCP precursor (pLHCP). When barley etiochloroplasts are incubated with this pLHCP, both labelled pLHCP and LHCP are found as integral thylakoid membrane proteins, incorporated into the major pigment-protein complex of the thylakoids. The presence of pLHCP in thylakoids and its proportion with respect to labelled LHCP depends on the developmental stage of the plastids used to study the import of pLHCP. The reduced amounts of chlorophyll in a chlorophyll b-less mutant of barley does not affect the proportion of pLHCP to LHCP found in the thylakoids when import of pLHCP into plastids isolated from the mutant plants is examined. Therefore, insufficient chlorophyll during early stages of plastid development does not seem to be responsible for their relative inefficiency in assembling pLHCP. A chase of labelled pLHCP that has been incorporated into the thylakoids of intact plastids, by further incubation of the plastids with unlabelled pLHCP, reveals that the pLHCP incorporated into the thylakoids can be processed to its mature size. Our observations strongly support the hypothesis that after import into plastids, pLHCP is inserted into thylakoids and then processed to its mature size under in vivo conditions.

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References

  1. Akoyunoglou G, Argyroudi-Akoyunoglou J: Post-translational regulation of chloroplast differentiation. In: Akoyunoglou G, Senger H (eds) Regulation of Chloroplast Differentiation, pp. 571–582. Alan R Liss, New York (1986).

    Google Scholar 

  2. 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 97: 183–188 (1979).

    Google Scholar 

  3. Apel K, Kloppstech K: The effect of light on the biosynthesis of the light-harvesting chlorophyll a/b protein. Evidence for the requirement of chlorophyll a for the stabilization of the apoprotein. Planta 150: 426–430 (1980).

    Google Scholar 

  4. Bellemare G, Bartlett SG, Chua N-H: Biosynthesis of chlorophyll a/b-binding polypeptides in wild type and the chlorina f2 mutant of barley. J Biol Chem 257: 7762–7767 (1982).

    Google Scholar 

  5. Bennett J: Biosynthesis of the light-harvesting chlorophyll a/b protein. Polypeptide turnover in darkness. Eur J Biochem 118: 61–70 (1981).

    Google Scholar 

  6. Chitnis PR, Harel E, Kohorn BD, Tobin EM, Thornber JP: Assembly of the precursor and processed light-harvesting chlorophyll a/b protein of Lemna into the light-harvesting complex II of barley etiochloroplasts. J Cell Biol 102: 982–988 (1986).

    Google Scholar 

  7. Chitnis PR, Nechushtai R, Harel E, Thornber JP: Some requirements for the insertion of the precursor of apoproteins of Lemna light-harvesting complex II into barley thylakoids. In: Biggins J (ed.) Progress in Photosynthesis Research, Vol. 4, pp. 573–576. Martinus Nijhoff, Dordrecht (1987).

    Google Scholar 

  8. Chitnis PR, Nechushtai R, Thornber JP: Insertion of the precursor of the light-harvesting chlorophyll a/b-protein into the thylakoids requires the presence of a developmentally regulated stromal factor. Plant Mol Biol 10: 3–12 (1987).

    Google Scholar 

  9. Chua N-H, Schmidt GW: Transport of proteins into mitochondria and chloroplasts. J Cell Biol 81: 461–483 (1979).

    Google Scholar 

  10. Cline K: Import of proteins into chloroplasts. Membrane integration of a thylakoid precursor protein reconstituted in chloroplast lysates. J Biol Chem 261: 14804–14810 (1986).

    Google Scholar 

  11. Cline K, Werner-Washburne M, Andrews J, Keegstra K: Thermolysin is a suitable protease for probing the surface of intact pea chloroplasts. Plant Physiol 75: 675–678 (1984).

    Google Scholar 

  12. Cline K, Werner-Washburne M, Lubben TH, Keegstra K: Precursors to two nuclear-encoded chloroplast proteins bind to the outer envelope membrane before being imported into chloroplasts. J Biol Chem 260: 3691–3696 (1985).

    Google Scholar 

  13. Gollmer I, Apel K: The phytochrome-controlled accumulation of mRNA sequences encoding the light-harvesting chlorophyll a/b protein of barley (Hordeum vulgare L.). Eur J Biochem 133: 309–313 (1983).

    Google Scholar 

  14. Grebanier AE, Coen DM, Rich A, Bogorad L: Membrane proteins synthesized but not processed by isolated maize chloroplasts. J Cell Biol 78: 734–746 (1978).

    Google Scholar 

  15. Hageman J, Robinson C, Smeekens S, Weisbeek P: A thylakoid processing protease is required for complete maturation of the lumen protein plastocyanin. Nature 324: 567–569 (1986).

    Google Scholar 

  16. Highkin HR: Chlorophyll studies on barley mutants. Plant Physiol 25: 294–306 (1950).

    Google Scholar 

  17. Karlin-Neumann GA, Tobin EM: Transit peptides of nuclear-encoded chloroplast proteins share a common amino-acid framework. EMBO J 5: 9–13 (1986).

    Google Scholar 

  18. Karlin-Neumann GA, Kohorn BD, Thornber JP, Tobin EM: A chlorophyll a/b-protein encoded by a gene containing an intron with characteristics of a transposable element. J Mol Appl Genet 3: 45–61 (1985).

    Google Scholar 

  19. Kessler SW: Use of protein A-bearing staphylococci for the immunoprecipitation and isolation of antigens from cells. Meth Enzymol 73: 442–446 (1981).

    Google Scholar 

  20. Kohorn BD, Harel E, Chitnis PR, Thornber JP, Tobin EM: Functional and mutational analysis of the light-harvesting chlorophyll a/b protein of thylakoid membranes. J Cell Biol 102: 972–981 (1986).

    Google Scholar 

  21. Kohorn BD, Tobin EM: Chloroplast import of light-harvesting chlorophyll a/b-proteins with different amino termini and transit peptides. Plant Physiol 82: 1172–1174 (1986).

    Google Scholar 

  22. Lamppa GK, Morelli G, Chua N-H: Structure and developmental regulation of a wheat gene encoding the major chlorophyll a/b-binding polypeptide. Mol Cell Biol 5: 1370–1378 (1985).

    Google Scholar 

  23. Leech RM: Chloroplast development in angiosperms: current knowledge and future prospects. In: Baker NR, Barber J (eds) Chloroplast Biogenesis, pp. 1–21. Elsevier Science Publishers, Amsterdam (1984).

    Google Scholar 

  24. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning — A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor; NY (1982).

    Google Scholar 

  25. Matsuoka M, Kano-Murakami Y, Yamamoto N: Nucleotide sequence of cDNA encoding the light-harvesting chlorophyll a/b-binding protein from maize. Nucleic Acids Res 15: 6302 (1987).

    Google Scholar 

  26. Mullet JE: The amino acid sequence of the polypeptide segment which regulates membrane adhesion (grana stacking) in chloroplasts. J Biol Chem 258: 9941–9948 (1983).

    Google Scholar 

  27. Mullet JE, Chua N-H: In vitro reconstitution of synthesis, uptake, and assembly of cytoplasmically synthesized chloroplast proteins. Methods Enzymol 97: 502–509 (1983).

    Google Scholar 

  28. Roberts BE, Paterson BM: Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc Natl Acad Sci USA 70: 2330–2334 (1973).

    Google Scholar 

  29. Robinson C, Ellis RJ: Transport of proteins into chloroplasts. The precursor of small subunit of ribulose bisphosphate carboxylase is processed to the mature size in two steps. Eur J Biochem 142: 343–346 (1984).

    Google Scholar 

  30. Schmidt GW, Bartlett SG, Grossman AR, Cashmore AR, Chua N-H: Biosynthetic pathways of two polypeptide subunits of the light-harvesting chlorophyll a/b protein complex. J Cell Biol 91: 468–478 (1981).

    Google Scholar 

  31. Schmidt GW, Mishkind ML. The transport of proteins into chloroplasts. Ann Rev Biochem 55: 879–912 (1986).

    Google Scholar 

  32. Stayton MM, Black M, Bedbrook J, Dunsmuir P: A novel chlorophyll a/b binding (Cab) protein gene from petunia which encodes the lower molecular weight Cab precursor protein. Nucl Acids Res 14: 9781–9797 (1986).

    Google Scholar 

  33. Steinback KE, Burke JJ, Arntzen CJ: Evidence for the role of surface-exposed segments of the light-harvesting complex in cation-mediated control of chloroplast structure and function. Arch Biochem Biophys 195: 546–557 (1979).

    Google Scholar 

  34. Thornber JP, Peter GF, Nechushtai R, Chitnis PR, Hunter FA, Tobin EM: Electrophoretic separation of chlorophyll-protein complexes and their apoproteins. In: Akoyunoglou G, Senger H (eds.) Regulation of Chloroplast Differentiation, pp. 249–258. Alan R. Liss, New York (1986).

    Google Scholar 

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Chitnis, P.R., Morishige, D.T., Nechushtai, R. et al. Assembly of the barley light-harvesting chlorophyll a/b proteins in barley etiochloroplasts involves processing of the precursor on thylakoids. Plant Mol Biol 11, 95–107 (1988). https://doi.org/10.1007/BF00015663

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