Altered replicative capacity of recombinant HIV type 1 containing mutations at codons 26 and 29 of the viral protease.
HIV-1 protease activity is essential for viral replication. In spite of the high rates of HIV mutation, the active site of protease (residues 24-29) is a conserved site, where mutations have not been previously described. To determine the effect of mutations at positions T26 and A29 of the viral protease and its viability in recombinant HIV-1 strains, Sup-t1 cells were transfected by electroporation with PCR products from a protease containing the 26X or 29X mutation. These mutations were constructed by mutagenesis site directed with degenerative primers. Both changes modified the replicative capacity of the virus: viruses containing the 26X mutation have delayed replication as compared to the control virus HTLVIII B; the presence of the 29X mutation in the viral protease results in the absence of HIV-1 replication. These findings confirm that this region of the viral protease appears to be necessary for the viability of HIV-1, and it could be a good target for antiviral therapy.
Melón
Santiago
S
Hospital Central de Asturias, Oviedo, Spain.
Gil-Roda
Cristina
C
de Oña
María
M
Erice
Alejo
A
D002460:Cell Line; D003062:Codon / Q000737:chemistry*; D016333:HIV Protease / Q000737:chemistry / Q000235:genetics*; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011995:Recombination, Genetic; D014779:Virus Replication
D003062:Codon; D016333:HIV Protease