Bovine leukemia virus protease: purification, chemical analysis, and in vitro processing of gag precursor polyproteins.
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.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D003062:Codon; D003545:Cysteine / Q000378:metabolism; D015683:Gene Products, gag; D001909:Leukemia Virus, Bovine / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000378:metabolism*; D012190:Retroviridae / Q000201:enzymology*; D012191:Retroviridae Proteins / Q000235:genetics*
D000596:Amino Acids; D003062:Codon; D015683:Gene Products, gag; D011498:Protein Precursors; D012191:Retroviridae Proteins; D010447:Peptide Hydrolases; D003545:Cysteine