New single-chain (type 1) ribosome-inactivating proteins (RIPs) were isolated from the seeds of Basella rubra L. (two proteins) and from the leaves of Bougainvillea spectabilis Willd. (one protein). These RIPs inhibit protein synthesis both in a cell-free system, with an IC50 (concentration causing 50% inhibition) in the 10-10 M range, and by various cell lines, with IC50s in the 10-8-10-6 M range. All three RIPs released adenine not only from rat liver ribosomes but also from Escherichia coli rRNA, polyadenylic acid, herring sperm DNA, and artichoke mottled crinkle virus (AMCV) genomic RNA, thus being polynucleotide:adenosine glycosidases. The proteins from Basella rubra had toxicity to mice similar to that of most type I RIPs (Barbieri et al., 1993, Biochim Biophys Acta 1154: 237-282) with an LD50 (concentration that is 50% lethal) <=8 mg·kg-1 body weight, whilst the RIP from Bougainvillea spectabilis had an LD50 >32 mg·kg-1. The N-terminal sequence of the two RIPs from Basella rubra had 80-93% identity, whereas it differed from the sequence of the RIP from Bougainvillea spectabilis. When tested with antibodies against various RIPs, the RIPs from Basella gave some cross-reactivity with sera against dianthin 32, and weak cross-reactivity with momordin I and momorcochin-S, whilst the RIP from Bougainvillea did not cross-react with any antiserum tested. An RIP from Basella rubra and one from Bougainvillea spectabilis were tested for antiviral activity, and both inhibited infection of Nicotiana benthamiana by AMCV.

Cited By :46Export Date: 16 July 2015CODEN: PLANACorrespondence Address: Stirpe, F.; Dipartimento di Patologia, Sperimentale dell’Univ. di Bologna, Via S. Giacomo 14, I-40126 Bologna, Italy; email: stirpef@alma.unibo.itChemicals/CAS: Antiviral Agents; DNA, 9007-49-2; momordin I (protein); N-Glycosyl Hydrolases, EC 3.2.2.-; Plant Proteins; Poly A, 24937-83-5; Protein Synthesis Inhibitors; RNA N-glycosidase, EC 3.2.2.22; RNA, Bacterial; RNA, ViralReferences: Anas, F.J., Rojo, M.A., Ferreras, J.M., Iglesias, R., Muǹoz, R., Rocher, A., Méndez, E., Girbés, T., Isolation and partial characterization of a new ribosome-inactivating protein from Petrocoptis glancifolia (Lag.) Boiss (1992) Planta, 186, pp. 532-540;Barbien, L., Stoppa, C., Bolognesi, A., Large scale chromatographic purification of ribosome-mactivating proteins (1987) J Chromatogr, 408, pp. 235-243; Barhieri, L., Ferreras, J.M., Barraco, A., Ricci, P., Stirpe, F., Some ribosome-inactivating proteins clepurinate ribosomal RNA a multiple sites (1992) Biochem J, 286, pp. 1-4; Barbieri, L., Battelli, M.O., Stirpe, F., Ribosome-mactivatia proteins from plants (1993) Biochim Biophys Ada, 1154, pp. 237-282; Barbieri, L., Gorini, P., Valbonesi, P., Castiglioni, P., Stirpe, F., Unexpected activity of saporins (1994) Nature, 372, p. 624; Barbieri, L., Valbonesi, P., Gorini, P., Pession, A., Stirpe, F., Polynucleotide:adenosine glycosidase activity of saporin-Ll effect on DNA. RNA and poly(A) (1996) Biochem J, 319, pp. 507-513; Barbieri, L., Valbonesi, P., Bonora, F., Gorini, P., Bolognesi, A., Stirpe, I., Polynucleotide:adenosine glycosidase activity of ribosome-inactivating proteins: Effect on DNA. RNA and poly(A) (1997) Nucleic Acids Res, 25, pp. 518-522; Battelli, M.G., Stirpe, F., Ribosome-inactivating proteins from plants (1995) Antivira Proteins in Higher Plants, pp. 39-64. , Chessin M, DeBorde D, Zipf A (eds). CRC Press. Boca Raton; Bolognesi, A., Olivieri, F., Battelli, M.G., Barbieri, L., Falasca, A.I., Parente, A., Del Vecchio Blanco, F., Stirpe, F., Rihosomeinactivating proteins (RNA N-glycosidases) from the seeds Suponuriu ocymoides and Vaccaria pyramidata (1995) Eur J Biochem, 228, pp. 935-940; Endo, Y., Tsurugi, K., RNA N-glycosidase activity of ricin Achain. Mechanism of action of the toxic lectin ricin on eukaryotic ribosomes (1987) J Biol Chem, 262, pp. 8128-8130; Ferreras, J.M., Barbieri, L., Girbés, T., Battelli, M.G., Rojo, M.A., Arias, F.J., Rocher, M.A., Stirpe, F., Distribution and properties of major ribosome-inactivatin proteins (28 S rRNA N-glycosidases) of the plant Saponaria off icinalis L (Caryophyllaceae) (1993) Biochim Biophys Acta, 1216, pp. 31-42; Habuka, N., Murakami, Y., Noma, M., Kudo, T., Horikoshi, K., Amino acid sequence of Mirahili. s antiviral protein, tola synthesis of its gene and expression in Escherichia coli (1989) J Bio Chem, 264, pp. 6629-6637; Llabuka, N., Akiyama, K., Tuge, H., Miyano, M., Matsumoto, T., Noma, M., Expression and secretion of Mirabilis antiviral protein in Exchcrichia coli and its inhibition of in vitro eukaryotic am prokaryotic protein synthesis (1990) J Biol Chem, 265, pp. 10988-10992; Hong, Y., Saunders, K., Hartley, M.R., Stanley, J., Resistance to gerninivirus infection by virus-induced expression of dianthin in transgenic plants (1996) Virology, 220, pp. 119-127; Irvin, J.D., Antiviral proteins from Phytolacca (1995) Antiviral Proteins in Higher Plants, pp. 65-94. , Chessin M. DeBorde D, Zipf A (eds). CRC Press. Boca Raton; Kubo, S., Ikeda, T., Imaizumi, S., Takanami, Y., Mikami, Y., A potent plant virus inhibitor found in Mirabilis jalapa L (1990) Ann Phytopath Soc Jpn, 56, pp. 481-487; Lam, Y.H., Wong, Y.S., Wang, B., Wong, R.N.S., Yeung, H.W., Shaw, P.C., Use of trichosanthin to reduce infection by turnip mosaic virus (1996) Plant Sci, 114, p. 117; Li, M.-X., Yeung, H.-W., Pan, L.-P., Chan, S.-I., Trichosanthin, a potent HIV-I inhibitor, can cleave supercoiled DNA in vitro (1991) Nucleic Acids Res, 22, pp. 6309-6312; Ling, J., Liu, W.-Y., Wang, T.P., Cleavage of supercoiled doublestranded DNA by several ribosome-inuctivating proteins in vitro (1994) FEBS Lett, 345, pp. 143-146; Lodge, J.K., Kamewski, W.K., Turner, N.F., Broad-spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein (1993) Proc Null Acad Sci USA, 90, pp. 7089-7093; Marchant, A., Hartley, M.R., The action of pokeweed antivira protein and ricin A-chain on mutants in the X-sarcin loop of Exchcrichia coli 23S ribosomal RNA (1995) J Mol Biol, 254, pp. 848-855; McCann, W.P., Hall, L.M., Siler, W., Barton, N., Whitley, R.J., High-pressure liquid Chromatographie methods for deterrninin arabinosyladenine-5'-monophosphatc. arabmosyladenine and arabinosylhypoxanthine in plasma and urine (1985) Antimicro Agents Chemother, 28, pp. 265-273; Montanaro, L., Sperti, S., Zamhoni, M., Denaro, M., Testoni, G., Gaspcri-Campani, A., Stirpc, F., Effects of modeccin on the steps of peptide chain elongation (1978) Biochem J, 176, pp. 371-379; Moore, G.E., Sandberg, A., Ulrich, K., Suspension cell culture and in vivo and in viiro chromosome constitution of mouse leukemia L1210 (1966) J Natl Cancer Inst, 35, pp. 405-421; Murihy, N.S., Nagarajan, K., Sastry, A.B., Effect of prophylactic sprays of leaf extracts on the infection of tobacco by tobacco mosaic virus (1981) Indian J Agric Sci, 51, pp. 792-795; Olivieri, F., Prasad, V., Valbonesi, P., Srivastava, S., Ghosal-Choudhury, P., Barbieri, L., Bolognesi, A., Stirpe, F., A systemic antiviral resistance-inducing protein isolated from Clcrodenilnim inerme Gaertn. is a polynucleotide:adenosine glycosidase Iribosome-inactivating protein (1996) FEBS Lett, 396, pp. 132-134; Orita, M., Nishikawa, F., Kohno, T., Mitsui, Y., Fndo, Y., Taira, K., Nishikawa, S., High-resolution NMR study of a GdAGA letranucleotide loop that is an improved substrate for ricin, a cytoioxic plant protein (1996) Nucleic Acids Res, 24, pp. 611-618; Parente, A., De Luca, P., Bolognesi, A., Barbieri, L., Battelli, M.G., Abbondanza, A., Sande, M.J.W., Stirpe, F., Purification and partial characterization of single-chain ribosome-inactivating proteins from the seeds of phytolacca dioica L (1993) Biochim Biophys Acta, 1216, pp. 43-49; Prasad, V., Srivastava, S., Varsha, Verma, H.N., Two basic proteins from Clerudendrum inerme Gaertn. are inducers of systemic antiviral resistance in susceptible plants (1995) Plant Sci, 110, pp. 73-82; Ramakrishnan, S., (1993) Cylotoxic Conjugates, , Landes. Austin; Roncuzzi, L., Gasperi-Campani, A., DNA-nuclease activity of the single-chain ribosome-inactivatmg proteins dianthin 30. saporin 6 and gelonin (1996) FEBS Lett, 392, pp. 16-20; Stirpe, F., Williams, D.O., Onyon, L.J., Legg, R.F., Stevens, W.A., Dianthins. ribosome-damaging proteins with anti-viral properties from Dianihux rarvophvllux L. (carnation) (1981) Biochem J, 195, pp. 399-405; Stirpe, F., Barbien, L., Gonni, P., Valbonesi, P., Bolognesi, A., Polito, L., Activities associated with the presence of ribosomeinactivating proteins increase in senescent and stressed leaves (1996) FEBS Lett, 382, pp. 309-312; Ushasri, V., Nagarajan, K., Reddy, T.S.N., Isolation and characterisation of inhibitor to tobacco mosaic virus from Basella alba L (1982) Tobacco Res, 8, pp. 39-43; Verma, H.N., Dwivedi, S.O., Properties of a virus inhibiting agent, isolated from plants which have been treated with leal extracts from Bougainvilleu speclahilis (1984) Physiol Plant Pathol, 25, pp. 93-101; Verma, H.N., Varsha, Baranwai, V.K., Kndogenous virus inhibitors from plants: Their physical and biological properties (1995) Antiviral Proteins in Higher Plants, pp. 1-21. , Chessin M. DeBorde D. Zipf A (eds). CRC Press. Boca Raton; Zamboni, M., Brigotti, M., Rambelli, F., Montanaro, L., Sperti, S., High-pressure-liquid-chromatographic and fluorimetric methods for the determination of adenine released from ribosomcs by ricin and gelonin (1989) Biochem J, 259, pp. 639-643