Transcriptional analysis of human herpesvirus-8 open reading frames 71, 72, 73, K14, and 74 in a primary effusion lymphoma cell line.
We examined the transcription and splicing of open reading frames (ORFs) 71 (K13)-74 of human herpesvirus-8 (HHV-8) in the primary effusion lymphoma cell line BCP-1 (latently infected with HHV-8), using a combination of NORTHERN blot analysis, RT-PCR, and rapid amplification of cDNA ends (PCR-RACE). The three genes encoded by ORFs 71, 72, and 73 [viral FLICE inhibitory protein (v-FLIP), v-cyclin, latent nuclear antigen (LNA)] are transcribed from a common transcription start site in BCP-1 cells uninduced (latent) or induced (lytic) with n-butyrate. The resulting transcript is spliced to yield a 5.32-kb message encoding LNA, v-cyclin, and v-FLIP and a 1.7-kb bicistronic message encoding v-cyclin and v-FLIP. The two genes encoded by ORFs K14 and 74 (v-Ox2 and v-GPCR) are transcribed as a 2.7-kb bicistronic transcript that is induced with n-butyrate. A small (149-bp) intron is spliced from the intragenic noncoding region immediately before the v-GPCR initiating codon. Examination of sequence elements in the promoter of the LNA/v-cyclin/v-FLIP operon revealed TAATGARAT and Octamer binding motifs characteristic of herpesvirus immediate-early genes. Sequence elements in the v-Ox2/v-GPCR promoter included AP1 and Zta-like (EBV Zebra transactivator) binding motifs consistent with the n-butyrate induction of this operon.
Talbot
S J
SJ
Department of Medical Microbiology, Medical School, The University of Edinburgh, Teviot Place, Edinburgh, EH8 9AG, United Kingdom. s.talbot@ed.ac.uk
Weiss
R A
RA
Kellam
P
P
Boshoff
C
C
D001483:Base Sequence; D015152:Blotting, Northern; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; D002352:Carrier Proteins / Q000235:genetics; D002874:Chromosome Mapping; D016213:Cyclins / Q000235:genetics; D019288:Herpesvirus 8, Human / Q000235:genetics*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008223:Lymphoma / Q000235:genetics / Q000821:virology*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics; D016366:Open Reading Frames; D010750:Phosphoproteins; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D014764:Viral Proteins
D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; C506386:CFLAR protein, human; D002352:Carrier Proteins; D016213:Cyclins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; D014764:Viral Proteins; C118485:latent nuclear antigen (LNA)