Unexpected patterns of Epstein-Barr virus transcription revealed by a high throughput PCR array for absolute quantification of viral mRNA

Virology. 2015 Jan 1;474:117-30. doi: 10.1016/j.virol.2014.10.030. Epub 2014 Nov 15.

Abstract

We have validated a flexible, high-throughput and relatively inexpensive RT-QPCR array platform for absolute quantification of Epstein-Barr virus transcripts in different latent and lytic infection states. Several novel observations are reported. First, during infection of normal B cells, Wp-initiated latent gene transcripts remain far more abundant following activation of the Cp promoter than was hitherto suspected. Second, EBNA1 transcript levels are remarkably low in all forms of latency, typically ranging from 1 to 10 transcripts per cell. EBNA3A, -3B and -3C transcripts are likewise very low in Latency III, typically at levels similar to or less than EBNA1 transcripts. Thirdly, a subset of lytic gene transcripts is detectable in Burkitt lymphoma lines at low levels, including: BILF1, which has oncogenic properties, and the poorly characterized LF1, LF2 and LF3 genes. Analysis of seven African BL biopsies confirmed this transcription profile but additionally revealed significant expression of LMP2 transcripts.

Keywords: Burkitt lymphoma; Epstein–Barr virus; HHV4; Herpesvirus; Latency; Lytic replication; Transcription; Tumor virus; Viral gene expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocytes / virology
  • Burkitt Lymphoma / virology
  • Cell Line, Tumor
  • Epstein-Barr Virus Nuclear Antigens / genetics
  • Gene Expression Regulation, Viral
  • Genes, Viral
  • Herpesvirus 4, Human / genetics*
  • Humans
  • RNA, Messenger / analysis*
  • RNA, Messenger / genetics*
  • RNA, Viral / analysis*
  • RNA, Viral / genetics*
  • Receptors, G-Protein-Coupled / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Transcription, Genetic
  • Viral Proteins / genetics
  • Virion / genetics
  • Virus Latency / genetics

Substances

  • BILF1 protein, Epstein-Barr virus
  • Epstein-Barr Virus Nuclear Antigens
  • RNA, Messenger
  • RNA, Viral
  • Receptors, G-Protein-Coupled
  • Viral Proteins
  • EBV-encoded nuclear antigen 1