Tryptophan 95, an amino acid residue of the Caprine arthritis encephalitis virus vif protein which is essential for virus replication.
The Caprine arthritis encephalitis virus (CAEV) vif gene was demonstrated to be essential for efficient virus replication. CAEV Vif deletion mutants demonstrated an attenuated replication phenotype in primary goat cell cultures and resulted in abortive infection when inoculated into goats. In this study, we determined the in vitro replication phenotype of five CAEV Vif point mutant infectious molecular clones and the ability of the corresponding in vitro translated Vif proteins to interact with the CAEV Pr55(gag) in the glutathione S--transferase (GST) binding assay. Here we show that (i) three of the mutants (S170E, S170G, S197G) behaved as the wild-type CAEV according to virus replication and Vif--Gag interactions; (ii) one mutant (Vif 6mut) was replication incompetent and bound weakly to GST-Gag fusion proteins; and (iii) one mutant (Vif RG) was impaired for replication while retaining its interaction properties. This mutant points out the critical importance of the CAEV Vif tryptophan residue at position 95 for efficient virus replication, defining for this lentivirus a functional domain unrelated to the Gag binding region.
Seroude
V
V
INSERM U372, BP 176, 13178 Marseille Cedex 09, France.
Audoly
G
G
Gluschankof
P
P
Suzan
M
M
D000595:Amino Acid Sequence; D000818:Animals; D016088:Arthritis-Encephalitis Virus, Caprine / Q000378:metabolism / Q000502:physiology*; D002478:Cells, Cultured; D015683:Gene Products, gag / Q000378:metabolism; D016346:Gene Products, vif / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006041:Goats; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011498:Protein Precursors / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D017386:Sequence Homology, Amino Acid; D014364:Tryptophan / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014779:Virus Replication / Q000502:physiology*
D015683:Gene Products, gag; D016346:Gene Products, vif; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D014364:Tryptophan; D005982:Glutathione Transferase