Characterization of the transcription map and Rev activity of a highly cytopathic feline immunodeficiency virus.
A highly cytopathic feline immunodeficiency virus, FIV-Oma, was previously isolated from a nondomestic cat. In this report, we describe experiments to characterize its transcription map and examine its Rev activity. The temporal progression of viral gene expression is similar to that of HIV-1. The splicing pattern of viral transcripts was determined by sequence analysis of RT-PCR-amplified viral cDNAs. In vitro transcription and translation of two putative rev cDNAs revealed that they encode at least one 22-kDa protein. The Rev-responsive element (RRE) of FIV-Oma, identified by computer-assisted RNA secondary structure analysis, was inserted into the intron of an HIV-1-derived reporter plasmid and used in a transient transfection assay for Rev activity. Cotransfection of the RRE construct with the two rev cDNA clones significantly increased the expression of the reporter gene linked to the RRE, indicating that both transcripts encode an active Rev protein. The Rev activity of FIV-Oma is 5 to 8 times higher than that of a domestic cat FIV isolate, FIV-PPR. Our experiments also demonstrate the heterologous interaction of FIV-PPR Rev with the FIV-Oma RRE, even though the RREs of the two viruses have very little nucleotide sequence identity.
Zou
L
L
Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
Barr
M C
MC
Hoose
W A
WA
Avery
R J
RJ
D000818:Animals; D001483:Base Sequence; D002415:Cats; D002874:Chromosome Mapping; D003588:Cytopathogenic Effect, Viral / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D015757:Genes, rev; D016089:Immunodeficiency Virus, Feline / Q000235:genetics* / Q000472:pathogenicity*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D014158:Transcription, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral