Transcriptional analysis of human herpesvirus-8 open reading frames 71, 72, 73, K14, and 74 in a primary effusion lymphoma cell line

Simon Talbot, R A Weiss, P Kellam, C Boshoff

Research output: Contribution to journalArticlepeer-review

Abstract

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.
Original languageEnglish
Pages (from-to)84-94
Number of pages11
JournalVirology
Volume257
Issue number1
DOIs
Publication statusPublished - 1999

Keywords

  • Base Sequence
  • Blotting, Northern
  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • Carrier Proteins
  • Chromosome Mapping
  • Cyclins
  • Herpesvirus 8, Human
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Lymphoma
  • Molecular Sequence Data
  • Nuclear Proteins
  • Open Reading Frames
  • Phosphoproteins
  • Polymerase Chain Reaction
  • Random Amplified Polymorphic DNA Technique
  • Transcription, Genetic
  • Tumor Cells, Cultured
  • Viral Proteins

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