Skip to main content

Advertisement

SpringerLink
Log in

Trichosanthin inhibits integration of human immunodeficiency virus type 1 through depurinating the long-terminal repeats

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Trichosanthin (TCS) is a type I ribosome-inactivating protein with potent inhibitory activity against human immunodeficiency virus type 1. However, the anti-viral mechanism remains elusive. By a well-accepted HIV-1 integration assay, we demonstrated that TCS prevents HIV-1 DNA integration in a dose dependent manner in cell culture. At the same condition, TCS fails to induce obvious cytotoxicity and is also unable to interference viral early events such as viral entry, uncoating or reverse transcription. The HIV-1 integrase can integrate HIV-1 long-terminal repeats into cellular chromosome. The interaction of TCS with these viral integration components was also examined, indicating that TCS does not interact with HIV-1 integrase by the GST-pull down assay, but binds to the long terminal repeats in a transient manner. We further revealed that TCS can efficiently depurinate HIV-1 long-terminal repeats, which may be responsible for the inhibitory activity on HIV-1 integration. In conclusion, we elucidated that TCS specifically inhibits HIV-1 integration by depurinating the long-terminal repeats.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Maraganore JM, Joseph M, Bailey MC (1987) Purification and characterization of trichosanthin. Homology to the ricin A chain and implications as to mechanism of abortifacient activity. J Biol Chem 262:11628–11633

    CAS  PubMed  Google Scholar 

  2. Zhang JS, Liu WY (1992) The mechanism of action of trichosanthin on eukaryotic ribosomes–RNA N-glycosidase activity of the cytotoxin. Nucleic Acids Res 20:1271–1275

    Article  CAS  PubMed  Google Scholar 

  3. McGrath MS, Hwang KM, Caldwell SE, Gaston I, Luk KC, Wu P, Ng VL, Crowe S, Daniels J, Marsh J et al (1989) 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 86:2844–2848

    Article  CAS  PubMed  Google Scholar 

  4. Byers VS, Levin AS, Waites LA, Starrett BA, Mayer RA, Clegg JA, Price MR, Robins RA, Delaney M, Baldwin RW (1990) A phase I/II study of trichosanthin treatment of HIV disease. AIDS 4:1189–1196

    Article  CAS  PubMed  Google Scholar 

  5. Byers VS, Levin AS, Malvino A, Waites L, Robins RA, Baldwin RW (1994) A phase II study of effect of addition of trichosanthin to zidovudine in patients with HIV disease and failing antiretroviral agents. AIDS Res Hum Retroviruses 10:413–420

    Article  CAS  PubMed  Google Scholar 

  6. Kahn JO, Gorelick KJ, Gatti G, Arri CJ, Lifson JD, Gambertoglio JG, Bostrom A, Williams R (1994) Safety, activity, and pharmacokinetics of GLQ223 in patients with AIDS and AIDS-related complex. Antimicrob Agents Chemother 38:260–267

    CAS  PubMed  Google Scholar 

  7. Kahn JO, Kaplan LD, Gambertoglio JG, Bredesen D, Arri CJ, Turin L, Kibort T, Williams RL, Lifson JD, Volberding PA (1990) The safety and pharmacokinetics of GLQ223 in subjects with AIDS and AIDS-related complex: a phase I study. AIDS 4:1197–1204

    Article  CAS  PubMed  Google Scholar 

  8. Parikh BA, Tumer NE (2004) Antiviral activity of ribosome inactivating proteins in medicine. Mini Rev Med Chem 4:523–543

    CAS  PubMed  Google Scholar 

  9. Wang JH, Nie HL, Tam SC, Huang H, Zheng YT (2002) Anti-HIV-1 property of trichosanthin correlates with its ribosome inactivating activity. FEBS Lett 531:295–298. doi:10.1016/S0014-5793(02)03539-1

    Article  CAS  PubMed  Google Scholar 

  10. Wang JH, Nie HL, Huang H, Tam SC, Zheng YT (2003) Independency of anti-HIV-1 activity from ribosome-inactivating activity of trichosanthin. Biochem Biophys Res Commun 302:89–94. doi:10.1016/S0006-291X(03)00119-0

    Article  CAS  PubMed  Google Scholar 

  11. Wang YY, Ouyang DY, Huang H, Chan H, Tam SC, Zheng YT (2005) Enhanced apoptotic action of trichosanthin in HIV-1 infected cells. Biochem Biophys Res Commun 331:1075–1080. doi:10.1016/j.bbrc.2005.03.230

    Article  CAS  PubMed  Google Scholar 

  12. Au TK, Collins RA, Lam TL, Ng TB, Fong WP, Wan DC (2000) The plant ribosome inactivating proteins luffin and saporin are potent inhibitors of HIV-1 integrase. FEBS Lett 471:169–172. doi:10.1016/S0014-5793(00)01389-2

    Article  CAS  PubMed  Google Scholar 

  13. Li MX, Yeung HW, Pan LP, Chan SI (1991) Trichosanthin, a potent HIV-1 inhibitor, can cleave supercoiled DNA in vitro. Nucleic Acids Res 19:6309–6312

    Article  CAS  PubMed  Google Scholar 

  14. Zhao J, Ben LH, Wu YL, Hu W, Ling K, Xin SM, Nie HL, Ma L, Pei G (1999) Anti-HIV agent trichosanthin enhances the capabilities of chemokines to stimulate chemotaxis and G protein activation, and this is mediated through interaction of trichosanthin and chemokine receptors. J Exp Med 190:101–111

    Article  CAS  PubMed  Google Scholar 

  15. Xia XF, Sui SF (2000) The membrane insertion of trichosanthin is membrane-surface-pH dependent. Biochem J 349(Pt 3):835–841

    CAS  PubMed  Google Scholar 

  16. Rosenbluh J, Hayouka Z, Loya S, Levin A, Armon-Omer A, Britan E, Hizi A, Kotler M, Friedler A, Loyter A (2007) Interaction between HIV-1 Rev and integrase proteins: a basis for the development of anti-HIV peptides. J Biol Chem 282:15743–15753

    Article  CAS  PubMed  Google Scholar 

  17. Lutzke RA, Vink C, Plasterk RH (1994) Characterization of the minimal DNA-binding domain of the HIV integrase protein. Nucleic Acids Res 22:4125–4131

    Article  CAS  PubMed  Google Scholar 

  18. Wang P, Tumer NE (1999) Pokeweed antiviral protein cleaves double-stranded supercoiled DNA using the same active site required to depurinate rRNA. Nucleic Acids Res 27:1900–1905

    Article  CAS  PubMed  Google Scholar 

  19. Barbieri L, Valbonesi P, Bonora E, Gorini P, Bolognesi A, Stirpe F (1997) Polynucleotide:adenosine glycosidase activity of ribosome-inactivating proteins: effect on DNA, RNA and poly(A). Nucleic Acids Res 25:518–522

    Article  CAS  PubMed  Google Scholar 

  20. Yamamoto N, Tanaka C, Wu Y, Chang MO, Inagaki Y, Saito Y, Naito T, Ogasawara H, Sekigawa I, Hayashida Y (2006) Analysis of human immunodeficiency virus type 1 integration by using a specific, sensitive and quantitative assay based on real-time polymerase chain reaction. Virus Genes 32:105–113. doi:10.1007/s11262-005-5851-2

    Article  CAS  PubMed  Google Scholar 

  21. Hayouka Z, Rosenbluh J, Levin A, Loya S, Lebendiker M, Veprintsev D, Kotler M, Hizi A, Loyter A, Friedler A (2007) Inhibiting HIV-1 integrase by shifting its oligomerization equilibrium. Proc Natl Acad Sci U S A 104:8316–8321. doi:10.1073/pnas.0700781104

    Article  CAS  PubMed  Google Scholar 

  22. Guiot E, Carayon K, Delelis O, Simon F, Tauc P, Zubin E, Gottikh M, Mouscadet JF, Brochon JC, Deprez E (2006) Relationship between the oligomeric status of HIV-1 integrase on DNA and enzymatic activity. J Biol Chem 281:22707–22719. doi:10.1074/jbc.M602198200

    Article  CAS  PubMed  Google Scholar 

  23. Lee-Huang S, Huang PL, Huang PL, Bourinbaiar AS, Chen HC, Kung HF (1995) Inhibition of the integrase of human immunodeficiency virus (HIV) type 1 by anti-HIV plant proteins MAP30 and GAP31. Proc Natl Acad Sci U S A 92:8818–8822

    Article  CAS  PubMed  Google Scholar 

  24. Barbieri L, Valbonesi P, Righi F, Zuccheri G, Monti F, Gorini P, Samori B, Stirpe F (2000) Polynucleotide:adenosine glycosidase is the sole activity of ribosome-inactivating proteins on DNA. J Biochem 128:883–889

    CAS  PubMed  Google Scholar 

  25. Kupfer PA, Leumann CJ (2007) The chemical stability of abasic RNA compared to abasic DNA. Nucleic Acids Res 35:58–68. doi:10.1093/nar/gkl948

    Article  PubMed  Google Scholar 

  26. Mazumder A, Pommier Y (1995) Processing of deoxyuridine mismatches and abasic sites by human immunodeficiency virus type-1 integrase. Nucleic Acids Res 23:2865–2871

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Dr. Jing-Yun Li for the help of HIV-1 work in Laboratory Biosafety Level-3 (BSL-3, Academy of Military Medical Sciences of China). We also thank Anna Cereseto for the plasmids expressing GST-IN and His-IN (International Centre for Genetic Engineering and Biotechnology, Trieste, Italy). This work was supported by National Natural Science Foundation of China and National Basic Research program of China (2004CB720005).

Author information

Authors and Affiliations

  1. State Key Laboratory of Biomembrane and Membrane Biotechnology, Department of Biological Sciences and Biotechnology, Tsinghua University, 100084, Beijing, People’s Republic of China

    Wen-Long Zhao, Du Feng, Ju Wu & Sen-Fang Sui

Authors
  1. Wen-Long Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Du Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ju Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sen-Fang Sui
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sen-Fang Sui.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhao, WL., Feng, D., Wu, J. et al. Trichosanthin inhibits integration of human immunodeficiency virus type 1 through depurinating the long-terminal repeats. Mol Biol Rep 37, 2093–2098 (2010). https://doi.org/10.1007/s11033-009-9668-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-009-9668-2

Keywords

Search

Navigation