Analysis of interferon-beta mRNA stability control after poly(I:C) stimulation using RNA metabolic labeling by ethynyluridine

Biochem Biophys Res Commun. 2012 Nov 9;428(1):44-9. doi: 10.1016/j.bbrc.2012.09.144. Epub 2012 Oct 9.

Abstract

Interferon-beta (IFN-β) is a critical antiviral cytokine and is essential for innate and acquired immune responses to pathogens. Treatment with polyinosinic:polycytidylic acid (poly(I:C)) induces transient accumulation of IFN-β mRNA, which involves an increase and a decrease of IFN-β mRNA. This phenomenon has been extensively analyzed as a model for understanding the mechanisms of transient gene induction in response to external stimuli. Using a new RNA metabolic labeling method with ethynyluridine to directly measure de novo RNA synthesis and RNA stability, we reassessed both de novo synthesis and degradation of IFN-β mRNA. We found that transcriptional activity is maintained after the maximum accumulation of IFN-β mRNA following poly(I:C) treatment on immortalized human bronchial epithelial cells. We also observed an unexpected change in the stability of IFN-β mRNA before and after the maximum accumulation. The results indicate that this method of RNA metabolic labeling provides a general approach for the simultaneous analysis of transcriptional activity and mRNA stability coupled with transcriptional timing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Humans
  • Interferon-beta / genetics*
  • Interferon-beta / immunology
  • Poly I-C / immunology
  • Poly I-C / pharmacology
  • RNA Stability*
  • RNA, Messenger / chemistry*
  • Transcription, Genetic*
  • Uridine / analogs & derivatives
  • Uridine / chemistry

Substances

  • 1-(3-C-ethynylribopentofuranosyl)uracil
  • RNA, Messenger
  • Interferon-beta
  • Poly I-C
  • Uridine