Characterization of the invariable residue 51 mutations of human immunodeficiency virus type 1 capsid protein on in vitro CA assembly and infectivity.
BACKGROUND
The mature HIV-1 conical core formation proceeds through highly regulated protease cleavage of the Gag precursor, which ultimately leads to substantial rearrangements of the capsid (CAp24) molecule involving both inter- and intra-molecular contacts of the CAp24 molecules. In this aspect, Asp51 which is located in the N-terminal domain of HIV-1 CAp24 plays an important role by forming a salt-bridge with the free imino terminus Pro1 following proteolytic cleavage and liberation of the CAp24 protein from the Pr55Gag precursor. Thus, previous substitution mutation of Asp51 to alanine (D51A) has shown to be lethal and that this invariable residue was found essential for tube formation in vitro, virus replication and virus capsid formation.


RESULTS
We extended the above investigation by introducing three different D51 substitution mutations (D51N, D51E, and D51Q) into both prokaryotic and eukaryotic expression systems and studied their effects on in vitro capsid assembly and virus infectivity. Two substitution mutations (D51E and D51N) had no substantial effect on in vitro capsid assembly, yet they impaired viral infectivity and particle production. In contrast, the D51Q mutant was defective both for in vitro capsid assembly and for virus replication in cell culture.


CONCLUSIONS
These results show that substitutions of D51 with glutamate, glutamine, or asparagine, three amino acid residues that are structurally related to aspartate, could partially rescue both in vitro capsid assembly and intra-cellular CAp24 production but not replication of the virus in cultured cells.
Abdurahman
Samir
S
Division of Clinical Virology, Karolinska Institutet, F68 Karolinska University Hospital, SE-141 86 Stockholm, Sweden. Samir.Abdurahman@ki.se
Youssefi
Masoud
M
Höglund
Stefan
S
Vahlne
Anders
A
D019943:Amino Acid Substitution; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D002478:Cells, Cultured; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D009154:Mutation; D019065:Virus Assembly / Q000502:physiology*; D014777:Virus Diseases / Q000235:genetics*; D014779:Virus Replication
D036022:Capsid Proteins