Search tips
Search criteria 


Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. 1988 October; 62(10): 3718–3728.
PMCID: PMC253515

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


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

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Atkins JF, Gesteland RF, Reid BR, Anderson CW. Normal tRNAs promote ribosomal frameshifting. Cell. 1979 Dec;18(4):1119–1131. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Cochran MA, 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. [PMC free article] [PubMed]
  • Debouck C, Gorniak JG, Strickler JE, Meek TD, Metcalf BW, 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. [PubMed]
  • Fox TD, 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. [PubMed]
  • Graham FL, van der Eb AJ. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. [PubMed]
  • Hanecak R, Semler BL, Ariga H, Anderson CW, 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. [PubMed]
  • 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. [PubMed]
  • Hinuma Y, Nagata K, Hanaoka M, Nakai M, Matsumoto T, Kinoshita KI, 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. [PubMed]
  • 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. [PubMed]
  • Hizi A, Henderson LE, Copeland TD, Sowder RC, Hixson CV, 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. [PubMed]
  • Iino T, Takeuchi K, Nam SH, 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. [PubMed]
  • 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. [PubMed]
  • Jacks T, Power MD, Masiarz FR, Luciw PA, Barr PJ, Varmus HE. Characterization of ribosomal frameshifting in HIV-1 gag-pol expression. Nature. 1988 Jan 21;331(6153):280–283. [PubMed]
  • Jacks T, Townsley K, Varmus HE, 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. [PubMed]
  • 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. [PubMed]
  • 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. [PubMed]
  • 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. [PubMed]
  • Kohno T, Roth JR. 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. [PubMed]
  • Kotler M, Katz RA, Skalka AM. Activity of avian retroviral protease expressed in Escherichia coli. J Virol. 1988 Aug;62(8):2696–2700. [PMC free article] [PubMed]
  • Kramer RA, Schaber MD, Skalka AM, Ganguly K, Wong-Staal F, Reddy EP. HTLV-III gag protein is processed in yeast cells by the virus pol-protease. Science. 1986 Mar 28;231(4745):1580–1584. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • 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. [PubMed]
  • Mous J, Heimer EP, Le Grice SF. Processing protease and reverse transcriptase from human immunodeficiency virus type I polyprotein in Escherichia coli. J Virol. 1988 Apr;62(4):1433–1436. [PMC free article] [PubMed]
  • Nam SH, 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. [PubMed]
  • Oroszlan S, Copeland TD. 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. [PubMed]
  • Palmenberg AC, Rueckert RR. Evidence for intramolecular self-cleavage of picornaviral replicase precursors. J Virol. 1982 Jan;41(1):244–249. [PMC free article] [PubMed]
  • Pearl LH, Taylor WR. Sequence specificity of retroviral proteases. Nature. 1987 Aug 6;328(6130):482–482. [PubMed]
  • Pearl LH, Taylor WR. A structural model for the retroviral proteases. Nature. 1987 Sep 24;329(6137):351–354. [PubMed]
  • Poiesz BJ, Ruscetti FW, Gazdar AF, Bunn PA, Minna JD, Gallo RC. 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. [PubMed]
  • Poiesz BJ, Ruscetti FW, Reitz MS, Kalyanaraman VS, Gallo RC. 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. [PubMed]
  • Ratner L, Josephs SF, Starcich B, Hahn B, Shaw GM, Gallo RC, 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. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Schwartz DE, Tizard R, Gilbert W. Nucleotide sequence of Rous sarcoma virus. Cell. 1983 Mar;32(3):853–869. [PubMed]
  • 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. [PubMed]
  • Shida H. Nucleotide sequence of the vaccinia virus hemagglutinin gene. Virology. 1986 Apr 30;150(2):451–462. [PubMed]
  • Shida H, Tochikura T, Sato T, Konno T, Hirayoshi K, Seki M, Ito Y, Hatanaka M, Hinuma Y, Sugimoto M, et al. Effect of the recombinant vaccinia viruses that express HTLV-I envelope gene on HTLV-I infection. EMBO J. 1987 Nov;6(11):3379–3384. [PubMed]
  • Strebel K, Beck E. A second protease of foot-and-mouth disease virus. J Virol. 1986 Jun;58(3):893–899. [PMC free article] [PubMed]
  • 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]
  • 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. [PubMed]
  • 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. [PubMed]
  • 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. [PubMed]
  • Venkatesan S, Baroudy BM, 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. [PubMed]
  • Vogt VM, 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. [PubMed]
  • 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. [PubMed]
  • 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. [PubMed]
  • 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. [PubMed]
  • Yoshinaka Y, Katoh I, Copeland TD, Oroszlan S. Translational readthrough of an amber termination codon during synthesis of feline leukemia virus protease. J Virol. 1985 Sep;55(3):870–873. [PMC free article] [PubMed]
  • Yoshinaka Y, Katoh I, Copeland TD, 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. [PubMed]
  • Yoshinaka Y, Katoh I, Copeland TD, Smythers GW, 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. [PMC free article] [PubMed]

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