title: Unexpected patterns of Epstein-Barr virus transcription revealed by a high throughput PCR array for absolute quantification of viral mRNA.
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.
authors.affiliation: School of Cancer Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
authors.forename: Rosemary J
authors.initials: RJ
authors.lastname: Tierney
authors.affiliation: School of Cancer Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
authors.forename: Claire D
authors.initials: CD
authors.lastname: Shannon-Lowe
authors.affiliation: School of Cancer Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
authors.forename: Leah
authors.initials: L
authors.lastname: Fitzsimmons
authors.affiliation: School of Cancer Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom. Electronic address: a.i.bell@bham.ac.uk.
authors.forename: Andrew I
authors.initials: AI
authors.lastname: Bell
authors.affiliation: School of Cancer Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
authors.forename: Martin
authors.initials: M
authors.lastname: Rowe
mesh_terms: D001402:B-Lymphocytes / Q000821:virology; D002051:Burkitt Lymphoma / Q000821:virology; D045744:Cell Line, Tumor; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics*; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics; D014771:Virion / Q000235:genetics; D017735:Virus Latency / Q000235:genetics
chemical_list: C495131:BILF1 protein, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; D012333:RNA, Messenger; D012367:RNA, Viral; D043562:Receptors, G-Protein-Coupled; D014764:Viral Proteins; C417029:EBV-encoded nuclear antigen 1
keywords: Burkitt lymphoma; Epstein–Barr virus; HHV4; Herpesvirus; Latency; Lytic replication; Transcription; Tumor virus; Viral gene expression