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Structural Analysis of Nuclear Genes Coding for the Precursor to the Small Subunit of Wheat Ribulose–1,5–Bisphosphate Carboxylase

Bio/Technology volume 1, pages 55–61 (1983)Cite this article

Abstract

We have isolated and characterized a full–length cDNA clone encoding the precursor to the small subunit of wheat ribulose–1,5–bisphosphate carboxylase. From the nucleotide sequence, we deduce that the precursor contains the mature small subunit of 128 amino acid residues and an amino–terminal transit sequence of 47 amino acid residues. Southern blot analysis reveals that the small subunit precursor is encoded by a multigene family. One member of the family has been isolated. Nucleotide sequence and R–loop analyses show that it contains only one intron which separates the coding sequence of the transit peptide from that of the mature protein. This gene is expressed in vivo, as shown by mRNA hybridization to a specific probe from its 3′ non–coding region.

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References

  1. Miziorko, H.M. & Lorimer, G.H. Ribulose-1,5-bisphosphate carboxylase-oxygenase. Ann Rev. Biochem. In press.

  2. Siegelman, H.W. & Hind, G. (eds) 1978. Photosynthetic carbon assimilation. Plenum Press, New York.

    Google Scholar 

  3. Coen, D.M., Bedbrook, J.R., Bogorad, L. & Rich, A. 1977. Maize chloroplast DNA fragment encoding the large subunit of ribulose bisphosphate carboxylase. Proc. Natl. Acad. Sci. U.S.A. 74, 5487–5491.

    Article  CAS  Google Scholar 

  4. Blair, G.E. & Ellis, R.J. 1973. Protein synthesis in chloroplasts. I. Light-driven synthesis of the large subunit of fraction I protein by isolated pea chloroplasts. Biochim. Biophys. Acta 319, 223–234.

    Article  CAS  Google Scholar 

  5. Kawashima, N. & Wildman, S.G. 1972. Studies on fraction I protein IV. Mode of inheritance of primary structure in relation to whether chloroplast or nuclear DNA contains the code for a chloroplast protein. Biochim. Biophys. Acta 262, 42–49.

    Article  CAS  Google Scholar 

  6. Highfield, P.E. & Ellis, R.J. 1978. Synthesis and transport of the small subunit of chloroplast ribulose bisphosphate carboxylase. Nature 271, 420–424.

    Article  CAS  Google Scholar 

  7. Chua, N.-H. & Schmidt, G.W. 1978. Post-translational transport into intact chloroplasts of a precursor to the small subunit of ribulose-1,5-bisphosphate carboxylase. Proc. Natl. Acad. Sci. U.S.A. 75, 6110–6114.

    Article  CAS  Google Scholar 

  8. Cashmore, A.R., Broadhurst, M.K. and Gray, R.E. 1978. Cell-free synthesis of leaf proteins: identification of an apparent precursor of the small subunit of ribulose-1,5-bisphosphate carboxylase. Proc. Natl. Acad. Sci. U.S.A. 75, 655–659.

    Article  CAS  Google Scholar 

  9. Link, G., Coen, D.M. & Bogorad, L. 1978. Differential expression of the gene for the large subunit of ribulose bisphosphate carboxylase in maize leaf cell types. Cell 15, 725–731.

    Article  CAS  Google Scholar 

  10. Broglie, R., Coruzzi, G. and Chua, N.-H. Differential expression of genes encoding polypeptides involved in C4 photosynthesis. In: Chloroplast Biogenesis Cambridge University Press, London (in press).

  11. Smith, S.M. & Ellis, R.J. 1978. Light-stimulated accumulation of transcripts of nuclear and chloroplast genes for ribulose-bisphosphate carboxylase. J. Mol. Appl. Genet. 1, 127–137.

    Google Scholar 

  12. Shinzaki, K., Sasaki, Y., Sakibama, T. & Kamikubo, T. 1982. Coordinate light-induction of two mRNAs, encoded in nuclei and chloroplasts of ribulose-1,5-bisphosphate carboxylase/oxygenase. FEES Lett. 144, 73–76.

    Article  Google Scholar 

  13. McIntosh, L., Poulsen, C. & Bogorad, L. 1980. Chloroplast gene sequence for the large subunit of ribulose bisphosphate carboxylase of maize. Nature 288, 556–560.

    Article  CAS  Google Scholar 

  14. Zurawski, G., Perrot, B., Bottomley, W. & Whitfeld, P. 1981. The structure of the gene for the large subunit of ribulose-1,5-bisphosphate carboxylase from spinach chloroplast DNA. Nucl. Acid Res. 14, 3251–3270.

    Article  Google Scholar 

  15. Broglie, R., Bellemare, G., Bartlett, S.G., Chua, N.-H. & Cashmore, A.R. 1981. Cloned DNA sequences complementary to mRNA encoding precursors to the small subunit of ribulose-1,5-bisphosphate carboxylase and a chlorophyll a/b binding polypeptide. Proc. Natl. Acad. Sci. U.S.A. 78, 7304–7308.

    Article  CAS  Google Scholar 

  16. Bedbrook, J.R., Smith, S.M. & Ellis, R.J. 1980. Molecular cloning and sequencing of cDNA encoding the precursor to the small subunit of chloroplast ribulose-1,5-bisphosphate carboxylase. Nature 287, 692–697.

    Article  CAS  Google Scholar 

  17. Coruzzi, G., Broglie, R., Cashmore, A.R. & Chua, N.-H. Nucleotide sequences of two pea cDNA clones encoding the small subunit of ribulose bisphosphate carboxylase and the major chlorophyll a/b-binding thylakoid polypeptide. J. Biol. Chem. 258, 1399–1402.

  18. Chua, N.-H., Grossman, A.R., Bartlett, S.G. & Schmidt, G.W. 1980. Synthesis, transport and assembly of chloroplast proteins. In: “Biological Chemistry of Organelle Formation” Th. Bucher, W. Sebald, and H. Weiss, (eds.) pp. 113–117 Springer-Verlag Press, New York.

    Chapter  Google Scholar 

  19. Bowman, C.M., Koller, B., Delius, H. & Dyer, T.A. 1981. A physical map of wheat chloroplast DNA showing the location of the structural genes for the ribosomal RNAs and the large subunit of ribulose-1,5-bisphosphate carboxylase. Mol. Gen. Genet. 183, 93–101.

    Article  CAS  Google Scholar 

  20. Thomas, P.S. 1980. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. U.S.A. 77, 5201–5205.

    Article  CAS  Google Scholar 

  21. Carmichael, G.G., McMaster, G.R. 1980. The analysis of nucleic acids in gels using glyoxal and acridine orange. In: Methods in enzymology, L. Grossman, K. Moldave (eds.) 65, 380–391. Academic Press, New York.

    Google Scholar 

  22. Southern, E.M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98, 503–517.

    Article  CAS  Google Scholar 

  23. Maniatis, T., Jeffrey, A. & Kleid, D.G. 1975. Nucleotide sequence of the rightward operator of phage lambda. Proc. Natl. Acad. Sci. U.S.A. 72, 1184–1188.

    Article  CAS  Google Scholar 

  24. Berry-Lowe, S.L., McKnight, T.D., Shah, D.M. & Meagher, R.B. 1982. The nucieotide sequence, expression and evolution of one member of a multigene family encoding the small subunit of ribulose-1,5-bisphosphate carboxylase in soybean. J. Mol. & Appl. Gen. 1, 483–498.

    CAS  Google Scholar 

  25. Rimm, D.L., Horness, D., Kucera, J. & Blattner, F.R. 1980. Construction of coliphage lambda charon vector with Bam HI cloning sites. Gene 12, 301–309.

    Article  CAS  Google Scholar 

  26. Scalenghe, F., Turco, E., Edstrom, J.E., Pirrotta, V. & Melli, M. 1981. Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes. Chromosoma (Berl.) 82, 205–216.

    Article  CAS  Google Scholar 

  27. Wellauer, P.K. & Dawid, I.B. 1977. The structural organization of ribosomal DNA in Drosophila melanogaster. Cell 10, 193–212.

    Article  CAS  Google Scholar 

  28. Maxam, A. & Gilbert, W. 1980. Sequencing end-labeled DNA with base-specific chemical cleavages. In: Methods in Enzymology L. Grossman, K. Moldave (eds.) Academic Press, New York. 18, 499–560.

    Google Scholar 

  29. Takruri, I.A.H., Boulter, D. & Ellis, R.J. 1981. Amino acid sequence of the small subunit of ribulose-1,5-bisphosphate carboxylase of Pisum sativum. Phytochem. 20, 413–415.

    Article  CAS  Google Scholar 

  30. Mount, S.M. 1982. A catalogue of splice junction sequences. Nucl. Acids Res. 10, 459–472.

    Article  CAS  Google Scholar 

  31. Douce, R. & Joyard, J. 1979. Structure and function of the plastid envelope. Adv. Bot. Res. 7, 1–116.

    CAS  Google Scholar 

  32. Coruzzi, G., Broglie, R., Lamppa, G. & Chua, N.-H. Expression of nuclear genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase. In: “Structure and Function of Plant Genomes” O. Ciferri, L. Dure, III (eds.) Plenum Press, (in press).

  33. Chou, P.Y. & Fasman, G.D. 1978. Empirical predictions of protein conformation. Ann. Rev. Biochem. 47, 251–276.

    Article  CAS  Google Scholar 

  34. Poulsen, C., Strobek, S. & Haslett, B. 1976. Studies on the primary structure of the small subunit of ribulose-1,5-bisphosphate carboxylase. In: Genetics and biogenesis of chloroplast and mitochondria Th. Bucher, et al. (ed.) Elsevier/ North Holland Biomedical Press, Amsterdam. 8817–8824.

    Google Scholar 

  35. Martin, P.G. 1979. Amino acid sequence of the small subunit of ribulose-1,5-bisphosphate carboxylase from spinach. Aust. J. Plant Physiol 6, 401–408.

    CAS  Google Scholar 

  36. Berk, A.J. & Sharp, P.A. 1977. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclase-digested hybrids. Cell 12, 721–732.

    Article  CAS  Google Scholar 

  37. Weaver, R.F. & Weissmann, C. 1979. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5′ termini of 15S Beta-globin mRNA precursor and mature 10S Beta-globin mRNA have identical map coordinates. Nucl. Acids Res. 7, 1175–1193.

    Article  CAS  Google Scholar 

  38. Sollner-Webb, B. & Reeder, R.H. 1979. The nucleotide sequence of the initiation and termination sites for ribosomal RNA transcription in X. laevis. Cell 18, 485–499.

    Article  CAS  Google Scholar 

  39. Breathnach, R. & Chambon, P. 1981. Organization and expression of eukaryotic split genes coding for proteins. Ann. Rev. Biochem. 50, 349–384.

    Article  CAS  Google Scholar 

  40. Vieira, J. & Messing, J. 1982. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19, 269–276.

    Article  Google Scholar 

  41. Quisenberry, K.S. & Reitz, L.P. (eds) 1967. Wheat and wheat improvement. The American Society of Agronomy, Inc., Madison, Wisconsin.

  42. Gilbert, W. 1979. Introns and exons: Playgrounds of evolution. In: Eukaryotic Gene Regulation. Symposium on Molecular and cellular biology. R. Axel, T. Maniatis, and F. Fox, (eds.) 14, 1–14 Academic Press, New York.

    Google Scholar 

  43. Lomedico, P., Rosenthal, N., Efstratiadis, A., Gilbert, W., Kolodner, R. & Tizard, R. 1979. The structure and evolution of the two nonallelic rat preproinsulin genes. Cell 18, 545–558.

    Article  CAS  Google Scholar 

  44. McKeown, M. & Firtel, R.A. 1981. Differential expression and 5′ end mapping of actin genes in Dictyostelium. Cell 24, 799–807.

    Article  CAS  Google Scholar 

  45. Tobin, S.L., Zulauf, E., Sanchez, F., Craig, E.A. & McCarthy, B.J. 1980. Multiple actin-related sequences in the Drosophila melanogaster genome. Cell 19, 121–131.

    Article  CAS  Google Scholar 

  46. Durica, D.S., Schloss, J.A. & Crain, W.R. 1980. Studies on the organization of actin gene sequences in the sea urchin: molecular cloning of an intron-containing sequence coding for a cytoplasmid actin. Proc. Nat. Acad. Sci. U.S.A. 77, 5683–5687.

    Article  CAS  Google Scholar 

  47. Andrews, J.T. & Abel, K.M. 1981. Kinetics and subunit interactions of ribulose bisphosphate carboxylase-oxygenase from the Cyanobacterium, Synechococcus sp. J. Biol. Chem. 256, 8445–8451.

    CAS  PubMed  Google Scholar 

  48. Gatenby, A.A., Castleton, J.A. & Saul, J.W. Expression in E. coli of the maize and wheat chloroplast genes for large subunit of ribulose bisphosphate carboxylase. Nature (London) 291, 117–121.

    Article  CAS  Google Scholar 

  49. Dean, C. & Leech, R.M. 1982. Genome expression during normal leaf development. II. Direct correlation between ribulose bisphosphate carboxylase content and nuclear ploidy in a polyploid series of wheat. Plant Physiol. 70, 1605–1608.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Laboratory of Plant Molecular Biology, The Rockefeller University, New York, New York, 10021

    Richard Broglie, Gloria Coruzzi, Gayle Lamppa, Brian Keith & Nam-Hai Chua

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  1. Richard Broglie
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Broglie, R., Coruzzi, G., Lamppa, G. et al. Structural Analysis of Nuclear Genes Coding for the Precursor to the Small Subunit of Wheat Ribulose–1,5–Bisphosphate Carboxylase. Nat Biotechnol 1, 55–61 (1983). https://doi.org/10.1038/nbt0383-55

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