Research Paper | Published:

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

Bio/Technologyvolume 1pages5561 (1983) | Download Citation



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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  1. 1

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

  2. 2

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

  3. 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.

  4. 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.

  5. 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.

  6. 6

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

  7. 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.

  8. 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.

  9. 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.

  10. 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. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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. 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.

  19. 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.

  20. 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.

  21. 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.

  22. 22

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

  23. 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.

  24. 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.

  25. 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.

  26. 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.

  27. 27

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

  28. 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.

  29. 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.

  30. 30

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

  31. 31

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

  32. 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. 33

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

  34. 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.

  35. 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.

  36. 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.

  37. 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.

  38. 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.

  39. 39

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

  40. 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.

  41. 41

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

  42. 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.

  43. 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.

  44. 44

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

  45. 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.

  46. 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.

  47. 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.

  48. 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.

  49. 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.

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  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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