Structure-function studies of the human immunodeficiency virus type 1 matrix protein, p17.
The human immunodeficiency virus type 1 (HIV-1) matrix protein, p17, plays important roles in both the early and late stages of the viral life cycle. Using our previously determined solution structure of p17, we have undertaken a rational mutagenesis program aimed at mapping structure-function relationships within the molecule. Amino acids hypothesized to be important for p17 function were mutated and examined for effect in an infectious proviral clone of HIV-1. In parallel, we analyzed by nuclear magnetic resonance spectroscopy the structure of recombinant p17 protein containing such substitutions. These analyses identified three classes of mutants that were defective in viral replication: (i) proteins containing substitutions at internal residues that grossly distorted the structure of recombinant p17 and prevented viral particle formation, (ii) mutations at putative p17 trimer interfaces that allowed correct folding of recombinant protein but produced virus that was defective in particle assembly, and (iii) substitution of basic residues in helix A that caused some relocation of virus assembly to intracellular locations and produced normally budded virions that were completely noninfectious.
Cannon
P M
PM
Department of Biochemistry, University of Oxford, United Kingdom.
Matthews
S
S
Clark
N
N
Byles
E D
ED
Iourin
O
O
Hockley
D J
DJ
Kingsman
S M
SM
Kingsman
A J
AJ
D000595:Amino Acid Sequence; D015683:Gene Products, gag / Q000737:chemistry / Q000502:physiology*; D015488:HIV Antigens / Q000737:chemistry / Q000502:physiology*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000737:chemistry* / Q000648:ultrastructure; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D013329:Structure-Activity Relationship; D014764:Viral Proteins; D014771:Virion / Q000648:ultrastructure; D014779:Virus Replication; D054301:gag Gene Products, Human Immunodeficiency Virus
D015683:Gene Products, gag; D015488:HIV Antigens; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C054858:p17 protein, Human Immunodeficiency Virus Type 1