title: Complete mutagenesis of the HIV-1 protease.
abstract: Retroviruses encode a protease which needs to be active for the production of infectious virions. A disabling mutation in the protease results in the production of non-infectious virus particles and examination of proteins from these mutant virions reveals unprocessed Gag and Gag-Pol precursor proteins, the substrates of the viral protease. Each amino acid of the HIV-1 protease was individually mutated using a simple mutagenesis procedure which is capable of introducing and identifying missense mutations in each residue of a protein. Phenotypic screening of these mutants in a heterologous assay system reveals three regions within the protease where multiple consecutive amino-acid residues are sensitive to mutation. These results show that random mutagenesis can be used to identify functionally important regions within a protein. Mutants with conditional phenotypes have also been identified within this collection.
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina, Chapel Hill 27599.
authors.forename: D D
authors.initials: DD
authors.lastname: Loeb
authors.forename: R
authors.initials: R
authors.lastname: Swanstrom
authors.forename: L
authors.initials: L
authors.lastname: Everitt
authors.forename: M
authors.initials: M
authors.lastname: Manchester
authors.forename: S E
authors.initials: SE
authors.lastname: Stamper
authors.forename: C A
authors.initials: CA
authors.lastname: Hutchison
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015683:Gene Products, gag; D016333:HIV Protease; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D011498:Protein Precursors / Q000378:metabolism; D012191:Retroviridae Proteins / Q000378:metabolism; D014169:Transformation, Bacterial
chemical_list: D015683:Gene Products, gag; D011498:Protein Precursors; D012191:Retroviridae Proteins; D010450:Endopeptidases; D016333:HIV Protease