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Import and processing of the precursor form of the gamma subunit of the chloroplast ATP synthase from tobacco

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Abstract

A cDNA clone encoding the complete precursor of the γ subunit of chloroplast ATP synthase has been isolated from a tobacco (Nicotiana tabacum) leaf cDNA library in λgt11. The 1.4 kb insert encodes a polypeptide of 377 amino acid residues, of which 55 residues constitute an N-terminal presequence and 322 residues make up the mature γ subunit. Hybridisation of the cDNA to Southern blots of tobacco genomic DNA indicates the presence of two genes in the haploid genome. Transcription and translation of the cDNA in vitro produced a protein of 41 kDa which was imported by isolated pea chloroplasts and processed to the mature 36 kDa subunit. The γ subunit precursor was processed to the mature size by a processing peptidase of 180 kDa present in pea stromal extracts.

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References

  1. Abad MS, Clark SE, Lamppa GK: Properties of a chloroplast enzyme that cleaves the chlorophyll a/b binding protein precursor. Plant Physiol 90: 17–124 (1989).

    Google Scholar 

  2. Bichler J, Herrmann RG: Analysis of the promoters of the single-copy genes for plastocyanin and subunit δ of the chloroplast ATP synthase from spinach. Eur J Biochem 190: 415–426 (1990).

    PubMed  Google Scholar 

  3. Deno H, Shinozaki K, Sugiura M: Nucleotide sequence of tobacco chloroplast gene for the α subunit of proton-translocating ATPase. Nucl Acids Res 11: 2185–2191 (1983).

    PubMed  Google Scholar 

  4. Feinberg AP, Vogelstein B: A technique for radiolabelling restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).

    PubMed  Google Scholar 

  5. Gray JC, Kung SD, Wildman SG, Sheen SJ: Origin of Nicotiana tabacum L. detected by polypeptide composition of Fraction I protein. Nature 252: 226–227 (1974).

    PubMed  Google Scholar 

  6. Inohara N, Iwamoto A, Moriyama Y, Shimomura S, Maeda M, Futai M: Two genes, atpC1 and atpC2, for the γ subunit of Arabidopsis thaliana chloroplast ATP synthase. J Biol Chem 266: 7333–7338 (1991).

    PubMed  Google Scholar 

  7. Iwamoto A, Miki J, Maeda M, Futai M: H+-ATPase γ subunit of Escherichia coli. Role of the conserved carboxyl-terminal region. J Biol Chem 265: 5043–5048 (1990).

    PubMed  Google Scholar 

  8. Ketcham SR, Davenport JW, Warncke K, McCarty RE: Role of the γ-subunit of chloroplast coupling factor 1 in the light dependent activation of photophosphorylation and ATPase activity by dithiothreitol. J Biol Chem 259: 7286–7293 (1984).

    PubMed  Google Scholar 

  9. Krebbers ET, Larrinua IM, McIntosh L, Bogorad L: The maize chloroplast genes for the β and ε subunits of the photosynthetic coupling factor CF1 are fused. Nucl Acids Res 10: 4985–5002 (1982).

    PubMed  Google Scholar 

  10. Mason JG, Whitfeld PR: The γ-subunit of spinach chloroplast ATP synthase: isolation and characterisation of cDNA and genomic clones. Plant Mol Biol 14: 1007–1018 (1990).

    PubMed  Google Scholar 

  11. Matsudaira P: Sequence from picomole quantities of protein electroblotted onto polyvinylidine difluoride membranes. J Biol Chem 262: 10035–10038 (1987).

    PubMed  Google Scholar 

  12. McCarty RE, Fagan J: Light-stimulated incorporation of N-ethylmaleimide into coupling factor 1 in spinach chloroplasts. Biochemistry 12: 1503–1507 (1973).

    PubMed  Google Scholar 

  13. Miki J, Maeda M, Mukohata Y, Futai M: The γ-subunit of ATP synthase from spinach chloroplasts. FEBS Lett 232: 221–226 (1988).

    Article  PubMed  Google Scholar 

  14. Mills JD, Mitchell P, Schürmann P: Modulation of coupling factor ATPase activity in intact chloroplasts. The role of the thioredoxin system. FEBS Lett 112: 173–177 (1980).

    Article  Google Scholar 

  15. Moroney JV, Fullmer CS, McCarty RE: Characterization of the cysteinyl-containing peptides of the γ-subunit of coupling factor 1. J Biol Chem 259: 7281–7285 (1984).

    PubMed  Google Scholar 

  16. Nalin CM, McCarty RE: Role of a disulfide bond in the γ-subunit in activation of the ATPase of chloroplast coupling factor 1. J Biol Chem 259: 7275–7280 (1984).

    PubMed  Google Scholar 

  17. Napier JA, Höglund A-S, Plant AL, Gray JC: Chloroplast import of the gamma subunit of ATP synthase. Plant Mol Biol, submitted.

  18. Nelson N: Proton-ATPase of chloroplasts. Curr Top Bioenerget 11: 1–33 (1981).

    Google Scholar 

  19. Newman BJ, Gray JC: Characterisation of a full-length cDNA clone for pea ferredoxin-NADP+ reductase. Plant Mol Biol 10: 511–520 (1988).

    Google Scholar 

  20. Pick U: Isolation of the ATPase complex (CF0-CF1). In: Edelman M, Hallick RB, Chua N-H (eds) Methods in Chloroplast Molecular Biology, pp. 873–880. Elsevier, Amsterdam, (1982).

    Google Scholar 

  21. Robinson C, Ellis RJ: Transport of proteins into chloroplasts. Partial purification of a chloroplast protease involved in the processing of imported precursor polypeptides. Eur J Biochem 142: 337–342 (1984).

    PubMed  Google Scholar 

  22. Shinohara K, Minami E-I, Watanabe A: Synthesis and assembly of H+-ATPase complex by isolated ‘rough’ thylakoids. Arch Biochem Biophys 260: 452–460 (1988).

    PubMed  Google Scholar 

  23. Tittgen J, Hermans J, Steppuhn J, Jansen T, Jansson C, Andersson B, Nechushtai R, Nelson N, Herrmann RG: Isolation of cDNA clones for fourteen nuclear-encoded thylakoid membrane proteins. Mol Gen Genet 204: 258–265 (1986).

    Google Scholar 

  24. Wales R, Newman BJ, Pappin D, Gray J: The extrinsic 33 kDa polypeptide of the oxygen-evolving complex of photosystem II is a putative calcium-binding protein and is encoded by a multi-gene family in pea. Plant Mol Biol 12: 439–451.

  25. Watanabe A, Price CA: Translation of mRNAs for subunits of chloroplast coupling factor 1 in spinach. Proc Natl Acad Sci USA 79: 6304–6308 (1982).

    Google Scholar 

  26. Westhoff P, Nelson N, Bünemann H, Herrmann RG: Localization of genes for coupling factor subunits on the spinach plastid chromosome. Curr Genet 4: 109–120 (1981).

    Google Scholar 

  27. Yu LM, Selman BR: cDNA sequence and predicted primary structure of γ-subunit from the ATP synthase from Chlamydomonas reinhardtii. J Biol Chem 263: 19342–19345 (1988).

    PubMed  Google Scholar 

  28. Zurawski G, Bottomley W, Whitfeld PR: Structure of the genes for the β and ε subunits of spinach chloroplast ATPase indicate a dicistronic mRNA and an overlapping translation stop/start signal. Proc Natl Acad Sci USA 79: 6260–6264 (1982).

    Google Scholar 

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Author notes

  1. K. H. Larsson

    Present address: Department of Cell Research, Swedish University of Agricultural Sciences, Box 7055, S-75007, Uppsala, Sweden

  2. J. A. Napier

    Present address: AFRC Institute of Arable Crops Research, Long Ashton Research Station, Long Ashton, BS18 9AF, Bristol, UK

Authors and Affiliations

  1. Department of Plant Sciences, University of Cambridge, Downing Street, CB2 3EA, Cambridge, UK

    K. H. Larsson, J. A. Napier & J. C. Gray

  2. Cambridge Centre for Molecular Recognition, University of Cambridge, Downing Street, CB2 3EA, Cambridge, UK

    K. H. Larsson, J. A. Napier & J. C. Gray

Authors
  1. K. H. Larsson
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  2. J. A. Napier
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  3. J. C. Gray
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Larsson, K.H., Napier, J.A. & Gray, J.C. Import and processing of the precursor form of the gamma subunit of the chloroplast ATP synthase from tobacco. Plant Mol Biol 19, 343–349 (1992). https://doi.org/10.1007/BF00027359

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  • DOI: https://doi.org/10.1007/BF00027359

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