Abstract
The ribosome-inactivating proteins (RIPs) are a family of plant enzymes for which a unique activity has been determined: rRNA N-glycosidase, which removes adenine at a specific universally conserved position (A4324 in the case of rat ribosomes). Here we report that saporin-L1, a RIP from the leaves of Saponaria officinalis, recognizes other substrates, including RNAs from different sources, DNA and poly(A). Saporin-L1 depurinated DNA extensively and released adenine from all adenine-containing polynucleotides tested. Adenine was the only base released from DNA or artificial polynucleotides. The characteristics of the reactions catalysed by saporin-L1 have been determined: optimal pH and temperature, ionic requirements, and the kinetic parameters Km and kcat. The reaction proceeded without cofactors, at low ionic strength, in the absence of Mg2+ and K+. Saporin-L1 had no activity towards various adenine-containing non-polynucleotide compounds (cytokinins, cofactors, nucleotides). This plant protein may now be classified as a polynucleotide: adenosine glycosidase.
Full Text
The Full Text of this article is available as a PDF (412.0 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- ALLEN E. H., SCHWEET R. S. Synthesis of hemoglobin in a cell-free system. I. Properties of the complete system. J Biol Chem. 1962 Mar;237:760–767. [PubMed] [Google Scholar]
- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barbieri L., Battelli M. G., Stirpe F. Ribosome-inactivating proteins from plants. Biochim Biophys Acta. 1993 Dec 21;1154(3-4):237–282. doi: 10.1016/0304-4157(93)90002-6. [DOI] [PubMed] [Google Scholar]
- Barbieri L., Ferreras J. M., Barraco A., Ricci P., Stirpe F. Some ribosome-inactivating proteins depurinate ribosomal RNA at multiple sites. Biochem J. 1992 Aug 15;286(Pt 1):1–4. doi: 10.1042/bj2860001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barbieri L., Gorini P., Valbonesi P., Castiglioni P., Stirpe F. Unexpected activity of saporins. Nature. 1994 Dec 15;372(6507):624–624. doi: 10.1038/372624a0. [DOI] [PubMed] [Google Scholar]
- Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
- Carnicelli D., Brigotti M., Montanaro L., Sperti S. Differential requirement of ATP and extra-ribosomal proteins for ribosome inactivation by eight RNA N-glycosidases. Biochem Biophys Res Commun. 1992 Jan 31;182(2):579–582. doi: 10.1016/0006-291x(92)91771-h. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Endo Y., Tsurugi K. The RNA N-glycosidase activity of ricin A-chain. The characteristics of the enzymatic activity of ricin A-chain with ribosomes and with rRNA. J Biol Chem. 1988 Jun 25;263(18):8735–8739. [PubMed] [Google Scholar]
- Ferreras J. M., Barbieri L., Girbés T., Battelli M. G., Rojo M. A., Arias F. J., Rocher M. A., Soriano F., Mendéz E., Stirpe F. Distribution and properties of major ribosome-inactivating proteins (28 S rRNA N-glycosidases) of the plant Saponaria officinalis L. (Caryophyllaceae). Biochim Biophys Acta. 1993 Oct 19;1216(1):31–42. doi: 10.1016/0167-4781(93)90034-b. [DOI] [PubMed] [Google Scholar]
- Girbés T., Barbieri L., Ferreras M., Arias F. J., Rojo M. A., Iglesias R., Alegre C., Escarmis C., Stirpe F. Effects of ribosome-inactivating proteins on Escherichia coli and Agrobacterium tumefaciens translation systems. J Bacteriol. 1993 Oct;175(20):6721–6724. doi: 10.1128/jb.175.20.6721-6724.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee-Huang S., Huang P. L., Kung H. F., Li B. Q., Huang P. L., Huang P., Huang H. I., Chen H. C. TAP 29: an anti-human immunodeficiency virus protein from Trichosanthes kirilowii that is nontoxic to intact cells. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6570–6574. doi: 10.1073/pnas.88.15.6570. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lindahl T. DNA repair enzymes. Annu Rev Biochem. 1982;51:61–87. doi: 10.1146/annurev.bi.51.070182.000425. [DOI] [PubMed] [Google Scholar]
- Marchant A., Hartley M. R. The action of pokeweed antiviral protein and ricin A-chain on mutants in the alpha-sarcin loop of Escherichia coli 23S ribosomal RNA. J Mol Biol. 1995 Dec 15;254(5):848–855. doi: 10.1006/jmbi.1995.0660. [DOI] [PubMed] [Google Scholar]
- McCann W. P., Hall L. M., Siler W., Barton N., Whitley R. J. High-pressure liquid chromatographic methods for determining arabinosyladenine-5'-monophosphate, arabinosyladenine, and arabinosylhypoxanthine in plasma and urine. Antimicrob Agents Chemother. 1985 Aug;28(2):265–273. doi: 10.1128/aac.28.2.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGrath M. S., Hwang K. M., Caldwell S. E., Gaston I., Luk K. C., Wu P., Ng V. L., Crowe S., Daniels J., Marsh J. GLQ223: an inhibitor of human immunodeficiency virus replication in acutely and chronically infected cells of lymphocyte and mononuclear phagocyte lineage. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2844–2848. doi: 10.1073/pnas.86.8.2844. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGrath M. S., Santulli S., Gaston I. Effects of GLQ223 on HIV replication in human monocyte/macrophages chronically infected in vitro with HIV. AIDS Res Hum Retroviruses. 1990 Aug;6(8):1039–1043. doi: 10.1089/aid.1990.6.1039. [DOI] [PubMed] [Google Scholar]
- Montanaro L., Sperti S., Zamboni M., Denaro M., Testoni G., Gasperi-Campani A., Stirpe F. Effect of modeccin on the steps of peptide-chain elongation. Biochem J. 1978 Nov 15;176(2):371–379. doi: 10.1042/bj1760371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakatsu K., Tasker R. A., Bukowskyj M. Shortening of liquid chromatography columns for reduced retention time. Anal Chem. 1983 Jul;55(8):1455–1456. doi: 10.1021/ac00259a069. [DOI] [PubMed] [Google Scholar]
- Olsen L. C., Aasland R., Wittwer C. U., Krokan H. E., Helland D. E. Molecular cloning of human uracil-DNA glycosylase, a highly conserved DNA repair enzyme. EMBO J. 1989 Oct;8(10):3121–3125. doi: 10.1002/j.1460-2075.1989.tb08464.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ready M. P., Katzin B. J., Robertus J. D. Ribosome-inhibiting proteins, retroviral reverse transcriptases, and RNase H share common structural elements. Proteins. 1988;3(1):53–59. doi: 10.1002/prot.340030105. [DOI] [PubMed] [Google Scholar]
- Stirpe F., Barbieri L., Gorini P., Valbonesi P., Bolognesi A., Polito L. Activities associated with the presence of ribosome-inactivating proteins increase in senescent and stressed leaves. FEBS Lett. 1996 Mar 18;382(3):309–312. doi: 10.1016/0014-5793(96)00188-3. [DOI] [PubMed] [Google Scholar]
- Stirpe F., Derenzini M., Barbieri L., Farabegoli F., Brown A. N., Knowles P. P., Thorpe P. E. Hepatotoxicity of immunotoxins made with saporin, a ribosome-inactivating protein from Saponaria officinalis. Virchows Arch B Cell Pathol Incl Mol Pathol. 1987;53(5):259–271. doi: 10.1007/BF02890252. [DOI] [PubMed] [Google Scholar]
- Teltow G. J., Irvin J. D., Aron G. M. Inhibition of herpes simplex virus DNA synthesis by pokeweed antiviral protein. Antimicrob Agents Chemother. 1983 Mar;23(3):390–396. doi: 10.1128/aac.23.3.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zamboni M., Brigotti M., Rambelli F., Montanaro L., Sperti S. High-pressure-liquid-chromatographic and fluorimetric methods for the determination of adenine released from ribosomes by ricin and gelonin. Biochem J. 1989 May 1;259(3):639–643. doi: 10.1042/bj2590639. [DOI] [PMC free article] [PubMed] [Google Scholar]