• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. Mar 1986; 57(3): 826–832.
PMCID: PMC252811

Bovine leukemia virus protease: purification, chemical analysis, and in vitro processing of gag precursor polyproteins.

Abstract

Bovine leukemia virus protease was purified to homogeneity and assayed by using murine leukemia virus Pr65gag, a polyprotein precursor of the viral core structural proteins, as the substrate. A chemical analysis of the protease, including an amino acid composition and NH2- and COOH-terminal amino acid sequence analysis, revealed that it has an Mr of 14,000 and is encoded by a segment of the viral RNA located between the gag gene and the putative reverse transcriptase gene. As expected from the nucleotide sequence data (Rice et al., Virology 142:357-377, 1985), the reading frame for the protease is different from both the gag and reverse transcriptase reading frames. The 5' end of the protease open reading frame extends 38 codons upstream from the codon for the NH2-terminal residue of the mature viral protease and overlaps the gag open reading frame by 7 codons. The 3' end of the protease open reading frame extends 26 codons beyond the codon for the COOH-terminal residue of the mature protease and overlaps 8 codons of the reverse transcriptase open reading frame. Several lines of evidence, such as protein mapping of the gag polyprotein precursor, the characteristic structure of the mRNA, and promotion of the synthesis of a gag polyprotein precursor by lysine tRNA in vitro, suggest that the protease could be translated by frameshift suppression of the gag termination codon. In vitro synthesized bovine leukemia virus gag-related polyproteins were cleaved by the protease into fragments which were the same size as the known components of bovine leukemia virus, suggesting that the specificity of cleavage catalyzed in vitro by the purified protease is the same as the specificity of cleavage found in the virus.

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 (1.9M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

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]
  • Chamberlain JP. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem. 1979 Sep 15;98(1):132–135. [PubMed]
  • Copeland TD, Morgan MA, Oroszlan S. Complete amino acid sequence of the nucleic acid-binding protein of bovine leukemia virus. FEBS Lett. 1983 May 30;156(1):37–40. [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]
  • Gazdar AF, Phillips LA, Sarma PS, Peebles PT, Chopra HC. Presence of sarcoma genome in a "non-infectious" mammalian virus. Nat New Biol. 1971 Nov 17;234(46):69–72. [PubMed]
  • Ghysdael J, Kettmann R, Burny A. Translation of bovine leukemia virus virion RNAs in heterologous protein-synthesizing systems. J Virol. 1979 Mar;29(3):1087–1098. [PMC free article] [PubMed]
  • Henderson LE, Copeland TD, Oroszlan S. Separation of amino acid phenylthiohydantoins by high-performance liquid chromatography on phenylalkyl support. Anal Biochem. 1980 Feb;102(1):1–7. [PubMed]
  • Hewick RM, Hunkapiller MW, Hood LE, Dreyer WJ. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem. 1981 Aug 10;256(15):7990–7997. [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]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [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]
  • Oroszlan S, Copeland TD, Henderson LE, Stephenson JR, Gilden RV. Amino-terminal sequence of bovine leukemia virus major internal protein: homology with mammalian type C virus p30 structural proteins. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2996–3000. [PMC free article] [PubMed]
  • Oroszlan S, Henderson LE, Stephenson JR, Copeland TD, Long CW, Ihle JN, Gilden RV. Amino- and carboxyl-terminal amino acid sequences of proteins coded by gag gene of murine leukemia virus. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1404–1408. [PMC free article] [PubMed]
  • Oroszlan S, Sarngadharan MG, Copeland TD, Kalyanaraman VS, Gilden RV, Gallo RC. Primary structure analysis of the major internal protein p24 of human type C T-cell leukemia virus. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1291–1294. [PMC free article] [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. [PMC free article] [PubMed]
  • Rice NR, Stephens RM, Burny A, Gilden RV. The gag and pol genes of bovine leukemia virus: nucleotide sequence and analysis. Virology. 1985 Apr 30;142(2):357–377. [PubMed]
  • Rice NR, Stephens RM, Couez D, Deschamps J, Kettmann R, Burny A, Gilden RV. The nucleotide sequence of the env gene and post-env region of bovine leukemia virus. Virology. 1984 Oct 15;138(1):82–93. [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. [PMC free article] [PubMed]
  • Schultz AM, Copeland TD, Oroszlan S. The envelope proteins of bovine leukemia virus: purification and sequence analysis. Virology. 1984 Jun;135(2):417–427. [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. [PMC free article] [PubMed]
  • Wray W, Boulikas T, Wray VP, Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981 Nov 15;118(1):197–203. [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. [PMC free article] [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. [PMC free article] [PubMed]
  • Yoshinaka Y, Luftig RB. Properties of a P70 proteolytic factor of murine leukemia viruses. Cell. 1977 Nov;12(3):709–719. [PubMed]
  • Yoshinaka Y, Luftig RB. p65 of Gazdar murine sarcoma viruses contains antigenic determinants from all four of the murine leukemia virus (MuLV) gag polypeptides (p15, p12, p30, and p10) and can be cleaved in vitro by the MuLV proteolytic activity. Virology. 1982 Apr 30;118(2):380–388. [PubMed]
  • Yoshinaka Y, Oroszlan S. Bovine leukemia virus post-envelope gene coded protein: evidence for expression in natural infection. Biochem Biophys Res Commun. 1985 Aug 30;131(1):347–354. [PubMed]

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

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...