Mature and functional viral miRNAs transcribed from novel RNA polymerase III promoters

Kevin W. Diebel, Anna L. Smith, Linda F. Van Dyk

Research output: Contribution to journalArticlepeer-review

59 Scopus citations

Abstract

Murid herpesvirus 4 (MuHV-4) microRNAs were previously cloned from latently infected tumor cells and predicted to be processed from a series of RNA polymerase III primary transcripts. We detected maturely processed MuHV-4 miRNAs within total RNA from lytically infected cells in vitro and infected tissues ex vivo, using a highly sensitive reverse ligation meditated RT-PCR strategy. We determined that the MuHV-4 microRNAs are biologically active during infection by a luciferase reporter system. We experimentally demonstrated that transcription of the MuHV-4 microRNAs is by RNA polymerase III by α-amanitin insensitivity and by specific deletion of the RNA polymerase III type 2-like promoter elements of MuHV-4, resulting in the complete loss of miRNA detection and function. Finally, we demonstrate that these 10 viral miRNAs, each transcribed from highly conserved and novel polymerase III promoter elements, vary markedly in their relative abundance and activity. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish (US)
Pages (from-to)170-185
Number of pages16
JournalRNA
Volume16
Issue number1
DOIs
StatePublished - Jan 2010

Keywords

  • Herpesvirus
  • MicroRNA
  • Noncoding RNA
  • RNA polymerase III
  • Reverse ligation mediated-reverse transcription-polymerase chain reaction

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