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Expression of Active Human Immunodeficiency Virus Reverse Transcriptase in Saccharomyces Cerevisiae

Bio/Technologyvolume 5pages486489 (1987) | Download Citation



The reverse transcriptase (RT) domain of the human immunodeficiency virus (HIV) pol gene was expressed in yeast. Purification and analysis of the recombinant protein indicated that yeast post–translational processing of RT was similar, if not identical, to that which had previously been observed in HIV infected lymphoid cells. In addition, recombinant RT was enzymatically active and could be inhibited by several anti–retroviral agents. These studies indicate that HIV RT expressed in yeast will be useful for the in vitro analysis of further potential inhibitors of this important enzyme.

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  1. 1

    Barré-Sinoussi, F., Chermann, J.C., Rey, F., Nugeyre, M.T., Chamaret, S., Gruest, J., Dauguet, C. and Axler-Blin, L. 1983. Isolation of a T-lymphotropic retrovirus from a patient at risk for Acquired Immune Deficiency Syndrome (AIDS). Science 220: 868–871.

  2. 2

    Wain-Hobson, S., Sonigo, P., Danos, O., Cole, S. and Alizon, M., 1985. Nucleotide Sequence of the AIDS Virus, LAV. Cell 40: 9–17.

  3. 3

    Popovic, M., Sarngadharan, M.G., Read, E. and Gallo, R.C., 1984. tection, isolation, and continuous production of cytopathic retrovir-uses (HTLV-III) from Patients with AIDS and Pre-AIDS. Science 224: 497–504.

  4. 4

    Ratner, L., et al. 1985. Complete nucleotide sequence of the AIDS virus, HTLV-III. Nature 313: 277–284.

  5. 5

    Levy, J.A., Hoffman, A.D., Kramer, S.M., Landis, J.A., Shimabu-kuro, J.M. and Oshiro, L.S., 1984. San Francisco patients with AIDS. Science 225: 840–842.

  6. 6

    Sanchez-Pescador, R., et al. 1985. Nucleotide sequence and expression of an AIDS-associated retrovirus (ARV-2). Science 227: 484–492.

  7. 7

    Baltimore, D. 1970. RNA-dependent DNA polymerase in virions of RNA tumor viruses. Nature 226: 1209–1211.

  8. 8

    Temin, H.M. and Mitzutani, S. 1970. RNA-directed DNA polymerase in virions of Rous sarcoma virus. Nature 226: 1211–1213.

  9. 9

    Veronese, F.M., Copeland, T.D., DeVico, A.L., Rahman, R., Orosz-lan, S., Gallo, R.C. and Sarngadharan, M.G. 1986. Characterization of highly immunogenic p66/p51 as the reverse transcriptase of HTLV-III/LAV. Science 231: 1289–1291.

  10. 10

    Mitsuya, H., Weinhold, K.J., Furman, P.A., St. Clair, M.H., Nusin-off, L., et al. 1985. 3′-Azido-3′-deoxythymidine (BW A509U): An antiviral agent that inhibits the infectivity and cytopathic effect of human T-lymphotropic virus type III/lymphadenopathy-associated virus in vitro. Proc. Natl. Acad. Sci. USA. 82: 7096–7100.

  11. 11

    Mitsuya, H. and Broder, S. 1986. Inhibition of the in vitro infectivity and cytopathic effect of human T-lymphotrophic virus type III/lymphadenopathy-associated virus (HTLV 11 I/LAV) by 2′, 3′-dideoxynu-cleosides. Proc. Natl. Acad. Sci. USA 83: 1911–1915.

  12. 12

    Luciw, P.A., Potter, S.J., Steimer, K., Dina, D. and Levy, J.A. 1984. Molecular cloning of AIDS-associated retrovirus. Nature 312: 760–763.

  13. 13

    Weiss, R., Teich, N., Varmus, H. and Coffin, J. 1982. RNA Tumor Viruses. Cold Spring Harbor Laboratory, New York.

  14. 14

    Tanese, N., Roth, M. and Goff, S.P. 1985. Expression of enzymatically active reverse transcriptase in Escherichia coli. Proc. Natl. Acad. Sci. USA 82: 4944–4948.

  15. 15

    Kotewicz, M.L., D'Alessio, M.D., Driftmier, K.M., Blodgett, K.P. and Gerard, G.F. 1985. Cloning and overexpression of Moloney murine leukemia virus reverse transcriptase in Escherichia coli. Gene 35: 249–258.

  16. 16

    Tanese, N., Sodroski, J., Haseltine, W.A. and Goff, S.P. 1986. Expression of reverse transcriptase activity of human T-lymphotropic virus Type III (HTLV III/LAV) in Escherichia coli. J. Virol. 59: 743–745.

  17. 17

    Travis, J., Owen, M., George, P., Carrell, R., Rosenberg, S., Hallewell, R.A. and Barr, P.J. 1985. Isolation and properties of recombinant DNA produced variants of human α1-proteinase inhibitor. J. Biol. Chem. 260: 4384–4389.

  18. 18

    Kramer, R.A., Schaber, M.D., Skalka, A.M., Ganguly, K., Wong-Staal, F. and Reddy, E.P. 1986. HTLV-III gag protein is processed in yeast Cells by the virus pol-protease. Science 231: 1580–1584.

  19. 19

    Hallewell, R.A., Mills, R., Tekamp-Olson, P., Blacher, R., Rosenberg, S., Otting, F., Masiarz, F.R. and Scandella, C.S., 1987. Human Cu, Zn superoxide dismutase produced in yeast: N-terminal acetylation and resistance to copper and zinc. Bio/Technology 5: 363–366.

  20. 20

    Takatsuji, H., Hirochika, H., Fukushi, T. and Ikeda, J.E. 1986. Expression of cauliflower mosaic virus reverse transcriptase in yeast. Nature 319: 240–243.

  21. 21

    Steimer, K.S., Puma, J.P., Power, M.D., Powers, M.A., George-Nascimento, C., et al. 1986. Differential antibody responses of individuals infected with AIDS-Associated retroviruses surveyed using the viral core antigen p25gag expressed in bacteria. Virology 150: 283–290.

  22. 22

    Lightfoote, M.M., Coligan, J.E., Folks, T.M., Fauci, A.S., Martin, M.A. and Venkatesan, S. 1986. Structural characterization of reverse transcriptase and endonuclease polypeptides of the acquired immunodeficiency syndrome retrovirus. J. Virol. 60: 771–775.

  23. 23

    Brake, A.J., Merryweather, J.P., Coit, D.G., Heberlein, U.A., Masiarz, F.R., et al. 1984. α-Factor-directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 81: 4642–4646.

  24. 24

    Barr, P.J., Parkes, D., Sabin, E.A., Power, M.D., Gibson, H.L., et al. 1986. Expression of functional domains of the AIDS associated retrovirus (ARV) in recombinant microorganisms. In: U.C.L.A. Symposium on Molecular and Cellular Biology, Vol. 43. Gallo, R.C., Haseltine, W., Klein, G., and Zur Hausen, H. (eds). Alan R. Liss, Inc. New York.

  25. 25

    Yarchoan, R., et al. 1986. Administration of 3 ′-azido-3′-deoxythymidine, an inhibitor of HTLV-III/LAV replication, to patients with AIDS or AIDS-related complex. Lancet (i): 575–580.

  26. 26

    Sandstrom, E.G., Kaplan, J.C., Byington, R.E. and Hirsch, M.S. 1985. Inhibition of human T Cell lymphotropic virus type III in vitro by phosphonoformate. Lancet (i): 1480–1482.

  27. 27

    Chermann, J.-C., Sinoussi, F.C. and Jasmin, C. 1975. Inhibition of RNA-dependent DNA polymerase of murine oncornaviruses by am-monium-5-tungsto-2-antimoniate. Biochem. Biophys. Res. Commun. 65: 1229–1236.

  28. 28

    Anand, R., Moore, J., Feorino, P., Curran, J. and Srinivasan, A., 1986. HTLV-III. Lancet (i): 97–98.

  29. 29

    Gurgo, C., Ray, R. and Green, M. 1972. Rifamycin derivatives strongly inhibiting RNA→DNA polymerase (reverse transcriptase) of murine sarcoma viruses. J. Natl. Cancer Inst. 49: 61–79.

  30. 30

    Grimley, P.M., Rosenblum, E.N., Mims, S.J. and Moss, B.J. 1970. Interruption by rifampin of an early stage in vaccinia virus morphoGenesis: Accumulation of membranes which are precursors of virus envelopes. Virology 6: 519–524.

  31. 31

    Maniatis, T., Fritsch, E.F. and Sambrook, J., 1982. Molecular Cloning, a Laboratory Manual. Cold Spring Harbor, New York.

  32. 32

    Messing, J. 1983. New M13 vectors for cloning. Methods Enzymol. 101: 20–78.

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  1. Philip J. Barr: Corresponding author.


  1. Chiron Corporation, 4560 Horton Street, Emeryville, CA, 94608

    • Philip J. Barr
    • , Michael D. Power
    • , Chun Ting Lee-Ng
    • , Helen L. Gibson
    •  & Paul A. Luciw


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