The caprine arthritis encephalitis virus tat gene is dispensable for efficient viral replication in vitro and in vivo.
Caprine arthritis encephalitis virus (CAEV) is a lentivirus closely related to visna virus and more distantly to other lentiviruses, such as human immunodeficiency virus. The genomes of visna virus and CAEV contain a tat gene encoding a protein able to weakly transactivate its own long terminal repeat, suggesting that transactivation may be a dispensable function for viral replication. Three different tat gene mutants of an infectious molecular clone of CAEV were used to study their replication after transfection or infection of primary goat synovial membrane cells and of blood-derived mononuclear cells or macrophages. Our results showed no difference between replication of the wild type and either the complete tat deletion mutant or the tat stop point mutant, whereas slower growth kinetics and lower levels of expression of the partial tat deletion mutant that of the wild type were obtained in these cells. Quantitative PCR and reverse transcription-PCR analyses of the different steps of a single replicative cycle revealed an identical pattern of retrotranscription, transcription, and viral production, whereas time course analysis demonstrated that the intracellular level of viral genomic RNA was affected by the partial tat deletion at later time points. We then compared the infectious properties of the wild-type and tat mutant viruses in vivo by direct inoculation of proviral DNAs into the joints of goats. All the animals seroconverted between 27 and 70 days postinoculation. Moreover, we were able to isolate tat mutant CAEV from blood-derived macrophages that was still able to infect synovial membrane cells in vitro. This study clearly demonstrates that the tat gene of CAEV is dispensable for viral replication in vitro and in vivo.
Harmache
A
A
Institut National de la Santé et de la Recherche Médicale, U372, Marseille, France.
Vitu
C
C
Russo
P
P
Bouyac
M
M
Hieblot
C
C
Peveri
P
P
Vigne
R
R
Suzan
M
M
D000818:Animals; D016088:Arthritis-Encephalitis Virus, Caprine / Q000235:genetics* / Q000502:physiology*; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D017353:Gene Deletion; D015696:Gene Products, tat / Q000032:analysis / Q000096:biosynthesis; D015756:Genes, tat; D016679:Genome, Viral; D006041:Goats; D006678:HIV / Q000235:genetics; D016180:Lentivirus Infections / Q000821:virology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D015183:Restriction Mapping; D015533:Transcriptional Activation; D014779:Virus Replication; D014790:Visna-maedi virus / Q000235:genetics; D054322:tat Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D015696:Gene Products, tat; D015345:Oligonucleotide Probes; D054322:tat Gene Products, Human Immunodeficiency Virus