Mutational analysis of human immunodeficiency virus type 1 protease suggests functional homology with aspartic proteinases.
Processing of the retroviral gag and pol gene products is mediated by a viral protease. Bacterial expression systems have been developed which permit genetic analysis of the human immunodeficiency virus type 1 protease as measured by cleavage of the pol protein precursor. Deletion analysis of the pol reading frame locates the sequences required to encode a protein with appropriate proteolytic activity near the left end of the pol reading frame but largely outside the gag-pol overlap region, which is at the extreme left end of pol. Most missense mutations within an 11-amino-acid domain highly conserved among retroviral proteases and with sequence similarity to the active site of aspartic proteinases abolish appropriate processing, suggesting that the retrovirus proteases share a catalytic mechanism with aspartic proteinases. Substitution of the amino acids flanking the scissile bond at three of the processing sites encoded by pol demonstrates distinct sequence requirements for cleavage at these different sites. The inclusion of a charged amino acid at the processing site blocks cleavage. A subset of these substitutions also inhibits processing at the nonmutated sites.
Loeb
D D
DD
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill 27599.
Hutchison
C A
CA
Edgell
M H
MH
Farmerie
W G
WG
Swanstrom
R
R
D000595:Amino Acid Sequence; D016282:Aspartic Acid Endopeptidases; D015153:Blotting, Western; D002872:Chromosome Deletion; D003062:Codon / Q000235:genetics; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D015683:Gene Products, gag; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010447:Peptide Hydrolases / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D012191:Retroviridae Proteins / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D003062:Codon; D015683:Gene Products, gag; D011498:Protein Precursors; D012191:Retroviridae Proteins; D010450:Endopeptidases; D010447:Peptide Hydrolases; D016282:Aspartic Acid Endopeptidases