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The lectin gene family of Ricinus communis: Cloning of a functional ricin gene and three lectin pseudogenes

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Abstract

Molecular hybridisation using a ricin cDNA probe has revealed that the ricin/Ricinus communis agglutinin (RCA) multigene family is composed of approximately eight members. Several genomic clones containing preproricin and preproricin-like sequences have been isolated. Partial analysis of three different genomic clones by DNA sequencing and ribonuclease protection has indicated that at least three members of the lectin gene family are non-functional. None of the original seventeen positive clones isolated appears to contain a Ricinus communis agglutinin (RCA) gene. One gene member analysed (pCBG3H1) represents a functional ricin gene similar in coding sequence to the published cDNA sequence and possesses typical eukaryotic consensus sequences and seed-specific elements within the flanking sequences. Investigation at the transcriptional level of the expression pattern of this gene revealed that mRNA accumulates during the post-testa stages of seed development. The pattern of accumulation of steady-state transcripts correlates closely with that previously observed at the protein and translatable RNA levels.

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References

  1. Araki T, Funatsu G: The complete amino acid sequence of the B chain of ricin E isolated from small grain castor bean seeds. Ricin E is a gene recombination product of ricin D and Ricinus communis agglutinin. Biochim Biophys Acta 911: 191–200 (1987).

    Google Scholar 

  2. Biggin MD, Gibson TJ, Hong GF: Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80: 3963–3965 (1983).

    Google Scholar 

  3. Dean C, Tamaki S, Dunsmuir P, Faureau M, Katayama C, Dooner H, Bedbrook J: mRNA transcripts of several plant genes are polyadenylated at multiple sites in vivo. Nucl Acids Res 14: 2229–2240 (1986).

    Google Scholar 

  4. Dickinson CD, Evans RP, Nielson NC: RY repeats are conserved on the 5′-flanking regions of legume seedprotein genes. Nucl Acids Res 16: 371 (1988).

    Google Scholar 

  5. Endo Y, Mitsui K, Motozuki M, Tsurugi K: The mechanism of action of ricin and related toxic lectins on eukaryotic ribosomes. J Biol Chem 262: 5908–5912 (1987).

    Google Scholar 

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

    Google Scholar 

  7. Funatsu G, Yoshitake S, Funatsu M: Primary structure of the lle chain of ricin D. Agric Biol Chem 42: 501–503 (1978).

    Google Scholar 

  8. Gallagher TF, Ellis RJ: Light-stimulated transcription of gene for two chloroplast polypeptides in isolated pea leaf nuclei. EMBO J 1: 1493–1498 (1982).

    Google Scholar 

  9. Gifford DJ, Greenwood JS, Bewley JD: Deposition of matrix and crystalloid storage proteins during protein body development in the endosperm of Ricinus communis L. cv. Hale seeds. Plant Physiol 69: 1471–1478 (1982).

    Google Scholar 

  10. Halling KC, Halling AC, Murray EE, Ladin BF, Houston LL, Weaver RF: Genomic cloning and characterisation of a ricin gene from Ricinus communis. Nucl Acids Res 13: 8019–8033 (1985).

    Google Scholar 

  11. Herrmann A, Schultz W, Hahlbrock K: Two alleles of the single-copy chalcone synthase gene in parsley differ by a transposon-like element. Mol Gen Genet 212: 93–98 (1988).

    Google Scholar 

  12. Irwin SD, Keen JN, Findlay JBC, Lord JM: The Ricinus communis 2S albumin precursor: A single preproprotein may be processed into two different heterodimeric storage protein. Mol Gen Genet 222: 400–408 (1990).

    Google Scholar 

  13. Kaminski PA, Buffard D, Strosberg AD: The pea lectin gene family contains only one functional gene. Plant Mol Biol 9: 497–507 (1987).

    Google Scholar 

  14. Keller GH, Taylor JM: Effect of hypophysectomy on the synthesis of rat liver albumin. J Biol Chem 251: 3768–3772 (1976).

    Google Scholar 

  15. Kislev N, Rubenstein I: Utility of ethidium bromide in the extraction from whole plants of high molecular weight maize DNA. Plant Physiol 66: 1140–1143 (1980).

    Google Scholar 

  16. Kreig PA, Melton DA. In vitro RNA synthesis with SP6 RNA polymerase. Meth Enzymol 155: 397–415 (1987).

    Google Scholar 

  17. ladin BF, Murray EE, Halling KC, Tilakaratne N, Long GL, Houston LL, Weaver RF. Characterisation of a cDNA encoding ricin E, a hybrid ricin-Ricinus communis agglutinin gene from the castor oil plant Ricinus communis. Plant Mol Biol 9: 287–295 (1987).

    Google Scholar 

  18. Lamb FI, Roberts LM, Lord JM: Nucleotide sequence of cDNA coding for preproricin. Eur J Biochem 148: 265–270 (1985).

    Google Scholar 

  19. Loenen WAM, Blattner FR: Lambda charon vectors (Ch32, 33, 34 and 35) adapted for DNA cloning in recombination-deficient hosts. Gene 26: 171–179 (1983).

    Google Scholar 

  20. Logemann J, Schell J, Willmitzer L: Improved method for the isolation of RNA from plant tissues. Anal Biochem 163: 16–20 (1987).

    Google Scholar 

  21. Lord JM, Roberts LM: Plant toxins. Microbiol Sci 4: 376–378 (1987).

    Google Scholar 

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

    Google Scholar 

  23. May MJ, Hartley MR, Roberts LM, Kreig PA, Osborn RW, Lord JM: Ribosome inactivation by ricin A chain: a sensitive method to assess the activity of wild-type and mutant polypeptides. EMBO J 8: 301–308 (1989).

    Google Scholar 

  24. Messing J, Vieira J: A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene 19: 269–276 (1982).

    Google Scholar 

  25. Olsnes S, Saltvedt E, Pihl A: Isolation and comparison of galactose-binding lectins from Abrus precatorius and Ricinus communis. J Biol Chem 249: 803–810 (1974).

    Google Scholar 

  26. Rigby PWJ, Dieckmann M, Rhodes C, Berg P: Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA Polymerase I. J Mol Biol 113: 237–251 (1977).

    Google Scholar 

  27. Roberts LM, Lamb FI, Lord JM: Biosynthesis and molecular cloning of ricin and RCA. 24. In: Membrane Mediated Cytotoxicity, pp. 73–82. Alan R. Liss, New York (1987).

    Google Scholar 

  28. Roberts LM, Lamb FI, Pappin DJC, Lord JM: The primary sequence of Ricinus communis agglutinin. Comparison with ricin. J Biol Chem 260: 15682–15686 (1985).

    Google Scholar 

  29. Roberts LM, Lord JM: Protein biosynthetic capacity in the endosperm tissue of ripening castor bean seeds. Planta 152: 420–427 (1981).

    Google Scholar 

  30. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  31. Vieira J, Messing J: The pUC plasmids, and M13 mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19: 259–268 (1982).

    Google Scholar 

  32. Vodkin LO, Rhodes PR, Goldberg RB: A lectin insertion has the structural features of a transposable element. Cell 34: 1023–1031 (1983).

    Google Scholar 

  33. Williams JG, Mason PJ: Hybridisation in the analysis of RNA. In: Glover DM (ed) DNA Cloning, vol. 2. IRL Press (1985).

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Tregear, J.W., Roberts, L.M. The lectin gene family of Ricinus communis: Cloning of a functional ricin gene and three lectin pseudogenes. Plant Mol Biol 18, 515–525 (1992). https://doi.org/10.1007/BF00040667

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  • DOI: https://doi.org/10.1007/BF00040667

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