Plant Molecular Biology

, Volume 22, Issue 1, pp 171–176 | Cite as

Nucleotide sequence of a genomic gene encoding tritin, a ribosome-inactivating protein from Triticum aestivum

  • Noriyuki Habuka
  • Jiro Kataoka
  • Masashi Miyano
  • Hideaki Tsuge
  • Hideo Ago
  • Masana Noma
Update section Short communication

Abstract

A genomic gene of tritin, a ribosome-inactivating protein (RIP) from Triticum aestivum, was cloned using a barley RIP gene as a probe. The 5′-non-coding region has potential TATA boxes and three sequences homologous to the binding sequence of the transcriptional activator protein Opaque-2 which activates maize RIP gene expression. The cloned DNA encoded tritin consits of 275 amino acids with no secretion signal sequence. The coding region of tritin was expressed in Escherichia coli using lac promoter and yielded a protein similar to the native one, as determined by SDS-polyacrulamide gel electrophoresis and immunological analysis.

Key words

expression genomic gene protein synthesis ribosome RNA N-glycosidase wheat germ 

Abbreviations

MAP

Mirabilis antiviral protein

RIP

ribosome-inactivating protein

SDS-PAGE

sodium dodecyl sulfate polyacrylamide gel electrophoresis

IC50

median of inhibitory concentration of L-[35S] methionine incorporation

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Asano K, Svensson B, Svendsen I, Poulsen FM: The complete primary structure of protein synthesis inhibitor II from barley seeds. Carlsberg Res Commun 51: 129–141 (1986).Google Scholar
  2. 2.
    Barbieri L, Stiipe F: Ribosome-inactivating proteins from plants: Properties and possible uses. Cancer Surv 1: 489–520 (1982).Google Scholar
  3. 3.
    Bass HW, Webster C, OBrian GR, Roberts JKM, Boston RS: A maize ribosome-inactivating protein is controlled by the transcriptional activator Opaque-2. Plant Cell 4: 225–234 (1992).Google Scholar
  4. 4.
    Coleman WH, Roberts WK: Inhibitors of animal cell-free protein synthesis from grains. Biochim Biophys Acta 696: 239–244 (1982).Google Scholar
  5. 5.
    Endo Y, Mitsui K, Motizuki M, Tsurugu K: The mechanism of action of ricin and related toxic lectines on eukaryotic ribosome. J Biol Chem 262: 5908–5912 (1987).Google Scholar
  6. 6.
    Endo Y, Tsurugi K: RNA N-glycosidase activity of ricin A-chain. J Biol Chem 262: 8128–8130 (1987).Google Scholar
  7. 7.
    Di Fonzo N, Manzocchi L, Salamini F, Soave C: Purification and properties of an endospermic protein of maize associated with the Opaque-2 and Opaque-6 genes. Planta 167: 587–594 (1986).Google Scholar
  8. 8.
    Habuka N, Murakami Y, Noma M, Kudo T, Horikoshi K: Amino acid sequence of Mirabilis antiviral protein, total synthesis of its gene and expression in Escherichia colt J Biol Chem 264: 6629–6637 (1989).Google Scholar
  9. 9.
    Habuka N, Akiyama K, Tsuge H, Miyano M, Matsumoto T, Noma M: Expression and secretion of Mirabilis antiviral protein in Escherichia coli and its inhibition of in vitro eukaryotic and prokaryotic protein synthesis. J Biol Chem 265: 10988–10992 (1990).Google Scholar
  10. 10.
    Habuka N, Miyano M, Kataoka J, Tsuge H, Noma M: Specificities of RNA N-glycosidase activity of Mirabilis antiviral protein variants. J Biol Chem 267: 7758–7760 (1992).Google Scholar
  11. 11.
    Halling KC, Halling AC, Murray EE, Ladin BF, Houston LL, Weaver RF: Genomic cloning and characterization of a ricin gene from Ricinus communis. Nucl Acids Res 13: 8019–8033 (1985).Google Scholar
  12. 12.
    Hartings H, Lazzaroni N, Marsan PA, Aragay A, Thompson R, Salamini F, Di Fonzo N, Palau J, Motto M: The b-32 protein from maize endosperm: characterization of genomic sequence encoding two alternative central domains. Plant Mol Biol 14: 1031–1040 (1990).Google Scholar
  13. 13.
    Kataoka J, Habuka N, Furuno M, Miyano M, Takanami Y, Koiwai M: DNA sequence of Mirabilis antiviral protein (MAP), a ribosome-inactivating protein with an antiviral property, from Mirabilis jarapa L. and expression in Escherichia coli. J Biol Chem 266: 8426–8430 (1991).Google Scholar
  14. 14.
    Kataoka J, Habuka N, Masuta C, Miyano M, Koiwai A: Isolation and analysis of a genomic clone encoding a pokeweed antiviral protein. Plant Mol Biol 20: 879–886 (1992).Google Scholar
  15. 15.
    Kataoka J, Habuka N, Miyano M, Masuta C, Koiwai A: Adenine depurination and inactivation of plant ribosomes by an antiviral protein of Mirabilis jarapa (MAP). Plant Mol Biol 20: 1111–1119 (1992).Google Scholar
  16. 16.
    Leah R, Tommerup H, Svendsen I, Mundy J: Biochemical and molecular characterization of three barley seed proteins with antifungal properties. J Biol Chem 266: 1564–1573 (1991).Google Scholar
  17. 17.
    Logemann L, Jach G, Tommerup H, Mundy J, Shell J: Expression of a barley ribosome-inactivating protein leads to increased fungal protection in transgenic tobacco plants. Bio/technology 10: 305–308 (1992).Google Scholar
  18. 18.
    Lohmer S, Maddaloni M, Motto M, Di Fonzo N, Hartings H, Salamini F, Thompson RD: The maize regulatory locus Opaque-2 encodes a DNA-binding protein which activates the transcription of the b-32 gene. EMBO J 10: 617–624 (1991).Google Scholar
  19. 19.
    Maki M, Takano E, Mori H, Sato A, Murachi T, Hatanala M: All four internally repetitive domains of pig calpastatin possess inhibitory activities against calpains I and II. FEBS Lett 1: 174–180 (1987).Google Scholar
  20. 20.
    McGrath MS, Hwang KM, Caldwell SE, Gaston I, Luk K-C, Wu P, Ng VL, Growe S, Daniels J, Marsh J, Deinhart T, Lekas PV, Vennari JC, Yeung H-W, Lifson JD: GLQ223: An inhibitor of human immunodeficiency virus replication in acutely and chronically infected cells of lymphocyte and mononuclear phagocyte lineage. Proc Natl Acad Sci USA 86: 2844–2848 (1989).Google Scholar
  21. 21.
    Ready MP, Brown DT, Robertus JD: Extracellular localization of pokeweed antiviral protein. Proc Natl Acad Sci USA 83: 5053–5056 (1986).Google Scholar
  22. 22.
    Reisbig RR, Bruland Ø: The protein synthesis inhibitors from wheat barley, and rye have identical antigenic determinants. Biochem Biophys Res Commun 114: 190–196 (1983).Google Scholar
  23. 23.
    Walsh TA, Morgan AE, Hey TD: Characterization and molecular cloning of a proenzyme form of a ribosome-inactivating protein from maize. J Biol Chem 266: 23422–23427 (1991).Google Scholar
  24. 24.
    Zarling JM, Moran PA, Haffar O, Sias J, Richman DD, Spina CA, Myers DE, Kuebelbeck V, Ledbetter JA, Uckun FM: Inhibition of HIV replication by pokeweed antiviral protein targeted to CD4+ cells by monoclonal antibodies. Nature 347: 92–95 (1990).Google Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Noriyuki Habuka
    • 1
  • Jiro Kataoka
    • 1
  • Masashi Miyano
    • 1
  • Hideaki Tsuge
    • 1
  • Hideo Ago
    • 1
  • Masana Noma
    • 1
  1. 1.Life Science Research LaboratoryJapan Tobacco, Inc.Yokohama, KanagawaJapan

Personalised recommendations