Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1988 Oct;62(10):3718–3728. doi: 10.1128/jvi.62.10.3718-3728.1988

Processing of gag precursor polyprotein of human T-cell leukemia virus type I by virus-encoded protease.

S H Nam 1, M Kidokoro 1, H Shida 1, M Hatanaka 1
PMCID: PMC253515  PMID: 2843670

Abstract

The biological activity encoded in the putative protease gene (pro) of human T-cell leukemia virus type I was investigated by using a vaccinia virus expression vector. The 53-kilodalton gag precursor polyprotein was processed into the mature p19, p24, and p15 gag proteins when the gag and protease-coding sequence was expressed under the control of a vaccinia virus promoter, suggesting that the protease may be synthesized through the mechanism of ribosomal frame shifting. The processing defect of a protease mutant could be complemented by cointroduction of a wild-type construct into the cell, demonstrating that the pro gene encodes the biologically active protease molecules which are capable of processing the gag precursor polyprotein in vivo in trans. A study involving the use of a variety of mutants constructed in vitro revealed that the protease consists of a nonessential carboxy-terminal region and a part essential for its activity, including the putative catalytic residue, aspartic acid. Furthermore, a cluster of adenine residues positioned at the overlapping region between the gag and pro genes was shown to be involved in the ribosomal frameshifting event for the synthesis of protease. To mimic the formation of the 76-kilodalton gag-pro precursor polyprotein formed by ribosomal slipping, the coding frames of the gag and pro gene were adjusted. The processing of the gag-pro precursor polyprotein depended on an intact protease gene, implying that a cis-acting function of human T-cell leukemia virus type I protease may be necessary to trigger the initial cleavage event that leads to the release of protease from the precursor protein.

Full text

PDF

Images in this article

3721Image
on p.3721
3722Image
on p.3722
3723Image
on p.3723
3723Image
on p.3723
3724Image
on p.3724
3725Image
on p.3725
3726Image
on p.3726

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atkins J. F., Gesteland R. F., Reid B. R., Anderson C. W. Normal tRNAs promote ribosomal frameshifting. Cell. 1979 Dec;18(4):1119–1131. doi: 10.1016/0092-8674(79)90225-3. [DOI] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cochran M. A., Puckett C., Moss B. In vitro mutagenesis of the promoter region for a vaccinia virus gene: evidence for tandem early and late regulatory signals. J Virol. 1985 Apr;54(1):30–37. doi: 10.1128/jvi.54.1.30-37.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Debouck C., Gorniak J. G., Strickler J. E., Meek T. D., Metcalf B. W., Rosenberg M. Human immunodeficiency virus protease expressed in Escherichia coli exhibits autoprocessing and specific maturation of the gag precursor. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8903–8906. doi: 10.1073/pnas.84.24.8903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fox T. D., Weiss-Brummer B. Leaky +1 and -1 frameshift mutations at the same site in a yeast mitochondrial gene. Nature. 1980 Nov 6;288(5786):60–63. doi: 10.1038/288060a0. [DOI] [PubMed] [Google Scholar]
  6. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  7. Hanecak R., Semler B. L., Ariga H., Anderson C. W., Wimmer E. Expression of a cloned gene segment of poliovirus in E. coli: evidence for autocatalytic production of the viral proteinase. Cell. 1984 Jul;37(3):1063–1073. doi: 10.1016/0092-8674(84)90441-0. [DOI] [PubMed] [Google Scholar]
  8. Hattori S., Kiyokawa T., Imagawa K., Shimizu F., Hashimura E., Seiki M., Yoshida M. Identification of gag and env gene products of human T-cell leukemia virus (HTLV). Virology. 1984 Jul 30;136(2):338–347. doi: 10.1016/0042-6822(84)90170-3. [DOI] [PubMed] [Google Scholar]
  9. Hinuma Y., Nagata K., Hanaoka M., Nakai M., Matsumoto T., Kinoshita K. I., Shirakawa S., Miyoshi I. Adult T-cell leukemia: antigen in an ATL cell line and detection of antibodies to the antigen in human sera. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6476–6480. doi: 10.1073/pnas.78.10.6476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hiramatsu K., Nishida J., Naito A., Yoshikura H. Molecular cloning of the closed circular provirus of human T cell leukaemia virus type I: a new open reading frame in the gag-pol region. J Gen Virol. 1987 Jan;68(Pt 1):213–218. doi: 10.1099/0022-1317-68-1-213. [DOI] [PubMed] [Google Scholar]
  11. Hizi A., Henderson L. E., Copeland T. D., Sowder R. C., Hixson C. V., Oroszlan S. Characterization of mouse mammary tumor virus gag-pro gene products and the ribosomal frameshift site by protein sequencing. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7041–7045. doi: 10.1073/pnas.84.20.7041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Iino T., Takeuchi K., Nam S. H., Siomi H., Sabe H., Kobayashi N., Hatanaka M. Structural analysis of p28 adult T-cell leukaemia-associated antigen. J Gen Virol. 1986 Jul;67(Pt 7):1373–1379. doi: 10.1099/0022-1317-67-7-1373. [DOI] [PubMed] [Google Scholar]
  13. Inoue J., Watanabe T., Sato M., Oda A., Toyoshima K., Yoshida M., Seiki M. Nucleotide sequence of the protease-coding region in an infectious DNA of simian retrovirus (STLV) of the HTLV-I family. Virology. 1986 Apr 15;150(1):187–195. doi: 10.1016/0042-6822(86)90278-3. [DOI] [PubMed] [Google Scholar]
  14. Jacks T., Power M. D., Masiarz F. R., Luciw P. A., Barr P. J., Varmus H. E. Characterization of ribosomal frameshifting in HIV-1 gag-pol expression. Nature. 1988 Jan 21;331(6153):280–283. doi: 10.1038/331280a0. [DOI] [PubMed] [Google Scholar]
  15. Jacks T., Townsley K., Varmus H. E., Majors J. Two efficient ribosomal frameshifting events are required for synthesis of mouse mammary tumor virus gag-related polyproteins. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4298–4302. doi: 10.1073/pnas.84.12.4298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Katoh I., Yasunaga T., Ikawa Y., Yoshinaka Y. Inhibition of retroviral protease activity by an aspartyl proteinase inhibitor. Nature. 1987 Oct 15;329(6140):654–656. doi: 10.1038/329654a0. [DOI] [PubMed] [Google Scholar]
  17. Katoh I., Yoshinaka Y., Rein A., Shibuya M., Odaka T., Oroszlan S. Murine leukemia virus maturation: protease region required for conversion from "immature" to "mature" core form and for virus infectivity. Virology. 1985 Sep;145(2):280–292. doi: 10.1016/0042-6822(85)90161-8. [DOI] [PubMed] [Google Scholar]
  18. Kobayashi N., Yamamoto N., Koyanagi Y., Schneider J., Hunsmann G., Hatanaka M. Translation of HTLV (human T-cell leukemia virus) RNA in a nuclease-treated rabbit reticulocyte system. EMBO J. 1984 Feb;3(2):321–325. doi: 10.1002/j.1460-2075.1984.tb01804.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kohno T., Roth J. R. A Salmonella frameshift suppressor that acts at runs of A residues in the messenger RNA. J Mol Biol. 1978 Nov 25;126(1):37–52. doi: 10.1016/0022-2836(78)90278-4. [DOI] [PubMed] [Google Scholar]
  20. Kotler M., Katz R. A., Skalka A. M. Activity of avian retroviral protease expressed in Escherichia coli. J Virol. 1988 Aug;62(8):2696–2700. doi: 10.1128/jvi.62.8.2696-2700.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kramer R. A., Schaber M. D., Skalka A. M., Ganguly K., Wong-Staal F., Reddy E. P. HTLV-III gag protein is processed in yeast cells by the virus pol-protease. Science. 1986 Mar 28;231(4745):1580–1584. doi: 10.1126/science.2420008. [DOI] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Mori K., Sabe H., Siomi H., Iino T., Tanaka A., Takeuchi K., Hirayoshi K., Hatanaka M. Expression of a provirus of human T cell leukaemia virus type I by DNA transfection. J Gen Virol. 1987 Feb;68(Pt 2):499–506. doi: 10.1099/0022-1317-68-2-499. [DOI] [PubMed] [Google Scholar]
  24. Mous J., Heimer E. P., Le Grice S. F. Processing protease and reverse transcriptase from human immunodeficiency virus type I polyprotein in Escherichia coli. J Virol. 1988 Apr;62(4):1433–1436. doi: 10.1128/jvi.62.4.1433-1436.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nam S. H., Hatanaka M. Identification of a protease gene of human T-cell leukemia virus type I (HTLV-I) and its structural comparison. Biochem Biophys Res Commun. 1986 Aug 29;139(1):129–135. doi: 10.1016/s0006-291x(86)80089-4. [DOI] [PubMed] [Google Scholar]
  26. Oroszlan S., Copeland T. D. Primary structure and processing of gag and env gene products of human T-cell leukemia viruses HTLV-ICR and HTLV-IATK. Curr Top Microbiol Immunol. 1985;115:221–233. doi: 10.1007/978-3-642-70113-9_14. [DOI] [PubMed] [Google Scholar]
  27. Palmenberg A. C., Rueckert R. R. Evidence for intramolecular self-cleavage of picornaviral replicase precursors. J Virol. 1982 Jan;41(1):244–249. doi: 10.1128/jvi.41.1.244-249.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pearl L. H., Taylor W. R. A structural model for the retroviral proteases. Nature. 1987 Sep 24;329(6137):351–354. doi: 10.1038/329351a0. [DOI] [PubMed] [Google Scholar]
  29. Pearl L. H., Taylor W. R. Sequence specificity of retroviral proteases. Nature. 1987 Aug 6;328(6130):482–482. doi: 10.1038/328482b0. [DOI] [PubMed] [Google Scholar]
  30. Poiesz B. J., Ruscetti F. W., Gazdar A. F., Bunn P. A., Minna J. D., Gallo R. C. Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7415–7419. doi: 10.1073/pnas.77.12.7415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Poiesz B. J., Ruscetti F. W., Reitz M. S., Kalyanaraman V. S., Gallo R. C. Isolation of a new type C retrovirus (HTLV) in primary uncultured cells of a patient with Sézary T-cell leukaemia. Nature. 1981 Nov 19;294(5838):268–271. doi: 10.1038/294268a0. [DOI] [PubMed] [Google Scholar]
  32. Ratner L., Josephs S. F., Starcich B., Hahn B., Shaw G. M., Gallo R. C., Wong-Staal F. Nucleotide sequence analysis of a variant human T-cell leukemia virus (HTLV-Ib) provirus with a deletion in pX-I. J Virol. 1985 Jun;54(3):781–790. doi: 10.1128/jvi.54.3.781-790.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sagata N., Yasunaga T., Tsuzuku-Kawamura J., Ohishi K., Ogawa Y., Ikawa Y. Complete nucleotide sequence of the genome of bovine leukemia virus: its evolutionary relationship to other retroviruses. Proc Natl Acad Sci U S A. 1985 Feb;82(3):677–681. doi: 10.1073/pnas.82.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schwartz D. E., Tizard R., Gilbert W. Nucleotide sequence of Rous sarcoma virus. Cell. 1983 Mar;32(3):853–869. doi: 10.1016/0092-8674(83)90071-5. [DOI] [PubMed] [Google Scholar]
  36. Seiki M., Hattori S., Hirayama Y., Yoshida M. Human adult T-cell leukemia virus: complete nucleotide sequence of the provirus genome integrated in leukemia cell DNA. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3618–3622. doi: 10.1073/pnas.80.12.3618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shida H. Nucleotide sequence of the vaccinia virus hemagglutinin gene. Virology. 1986 Apr 30;150(2):451–462. doi: 10.1016/0042-6822(86)90309-0. [DOI] [PubMed] [Google Scholar]
  38. Shida H., Tochikura T., Sato T., Konno T., Hirayoshi K., Seki M., Ito Y., Hatanaka M., Hinuma Y., Sugimoto M. Effect of the recombinant vaccinia viruses that express HTLV-I envelope gene on HTLV-I infection. EMBO J. 1987 Nov;6(11):3379–3384. doi: 10.1002/j.1460-2075.1987.tb02660.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Strebel K., Beck E. A second protease of foot-and-mouth disease virus. J Virol. 1986 Jun;58(3):893–899. doi: 10.1128/jvi.58.3.893-899.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tanaka Y., Koyanagi Y., Chosa T., Yamamoto N., Hinuma Y. Monoclonal antibody reactive with both p28 and p19 of adult T-cell leukemia virus-specific polypeptides. Gan. 1983 Jun;74(3):327–330. [PubMed] [Google Scholar]
  41. Tanaka Y., Lee B., Inoi T., Tozawa H., Yamamoto N., Hinuma Y. Antigens related to three core proteins of HTLV-I (p24, p19 and p15) and their intracellular localizations, as defined by monoclonal antibodies. Int J Cancer. 1986 Jan 15;37(1):35–42. doi: 10.1002/ijc.2910370107. [DOI] [PubMed] [Google Scholar]
  42. Toh H., Kikuno R., Hayashida H., Miyata T., Kugimiya W., Inouye S., Yuki S., Saigo K. Close structural resemblance between putative polymerase of a Drosophila transposable genetic element 17.6 and pol gene product of Moloney murine leukaemia virus. EMBO J. 1985 May;4(5):1267–1272. doi: 10.1002/j.1460-2075.1985.tb03771.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Venkatesan S., Baroudy B. M., Moss B. Distinctive nucleotide sequences adjacent to multiple initiation and termination sites of an early vaccinia virus gene. Cell. 1981 Sep;25(3):805–813. doi: 10.1016/0092-8674(81)90188-4. [DOI] [PubMed] [Google Scholar]
  45. Vogt V. M., Eisenman R., Diggelmann H. Generation of avian myeloblastosis virus structural proteins by proteolytic cleavage of a precursor polypeptide. J Mol Biol. 1975 Aug 15;96(3):471–493. doi: 10.1016/0022-2836(75)90174-6. [DOI] [PubMed] [Google Scholar]
  46. Yasunaga T., Sagata N., Ikawa Y. Protease gene structure and env gene variability of the AIDS virus. FEBS Lett. 1986 Apr 21;199(2):145–150. doi: 10.1016/0014-5793(86)80468-9. [DOI] [PubMed] [Google Scholar]
  47. Yoshida M., Miyoshi I., Hinuma Y. Isolation and characterization of retrovirus from cell lines of human adult T-cell leukemia and its implication in the disease. Proc Natl Acad Sci U S A. 1982 Mar;79(6):2031–2035. doi: 10.1073/pnas.79.6.2031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Yoshinaka Y., Katoh I., Copeland T. D., Oroszlan S. Murine leukemia virus protease is encoded by the gag-pol gene and is synthesized through suppression of an amber termination codon. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1618–1622. doi: 10.1073/pnas.82.6.1618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yoshinaka Y., Katoh I., Copeland T. D., Oroszlan S. Translational readthrough of an amber termination codon during synthesis of feline leukemia virus protease. J Virol. 1985 Sep;55(3):870–873. doi: 10.1128/jvi.55.3.870-873.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Yoshinaka Y., Katoh I., Copeland T. D., Smythers G. W., Oroszlan S. Bovine leukemia virus protease: purification, chemical analysis, and in vitro processing of gag precursor polyproteins. J Virol. 1986 Mar;57(3):826–832. doi: 10.1128/jvi.57.3.826-832.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. von der Helm K. Cleavage of Rous sarcoma viral polypeptide precursor into internal structural proteins in vitro involves viral protein p15. Proc Natl Acad Sci U S A. 1977 Mar;74(3):911–915. doi: 10.1073/pnas.74.3.911. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES