Equine infectious anemia virus gene expression: characterization of the RNA splicing pattern and the protein products encoded by open reading frames S1 and S2.
The utilization of predicted splice donor and acceptor sites in generating equine infectious anemia virus (EIAV) transcripts in fetal donkey dermal cells (FDD) was examined. A single splice donor site identified immediately upstream of the gag coding region joins the viral leader sequence to all downstream exons of spliced EIAV transcripts. The predominant 3.5-kb transcript synthesized in EIAV-infected FDD cells appears to be generated by a single splicing event which links the leader sequence to the first of two functional splice acceptor sites near the 5' end of the S1 open reading frame (ORF). The translation products encoded by the 3.5-kb transcript were examined by producing in vitro transcripts from a cDNA corresponding to this RNA followed by in vitro translation in wheat germ extracts. These transcripts directed the synthesis of three proteins: the virus trans-activator protein (EIAV Tat) encoded by ORF S1, a protein of unknown function encoded by ORF S2, and the virus envelope glycoprotein. When transfected into FDD cells, this cDNA also directed expression of EIAV Tat. Amino-terminal sequence analysis of the in vitro-synthesized S1 protein supports the suggestion that translation of EIAV Tat is initiated at a CUG codon within the virus leader region. Both in vitro-synthesized S2 protein and synthetic peptides corresponding to S2 are shown to react positively with sera obtained from EIAV-infected horses, providing the first direct evidence of expression of this protein in infected animals.
Schiltz
R L
RL
Department of Biochemistry, Louisiana State University, Baton Rouge 70803.
Shih
D S
DS
Rasty
S
S
Montelaro
R C
RC
Rushlow
K E
KE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015967:Gene Expression Regulation, Viral; D004860:Infectious Anemia Virus, Equine / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010527:Perissodactyla; D012322:RNA Precursors / Q000235:genetics; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014759:Viral Envelope Proteins / Q000096:biosynthesis*
D015345:Oligonucleotide Probes; D012322:RNA Precursors; D012333:RNA, Messenger; D014759:Viral Envelope Proteins; D015719:Single-Strand Specific DNA and RNA Endonucleases