Analysis of interferon-beta mRNA stability control after poly(I:C) stimulation using RNA metabolic labeling by ethynyluridine.
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.
Department of Cardiology, Yokohama City University School of Medicine, 3-9, Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan. kaitoabe@yokohama-cu.ac.jp
Kaito
K
Abe
Tomoaki
T
Ishigami
Ann-Bin
AB
Shyu
Shigeo
S
Ohno
Satoshi
S
Umemura
Akio
A
Yamashita
D002460:Cell Line; D006801:Humans; D016899:Interferon-beta / Q000235:genetics* / Q000276:immunology; D011070:Poly I-C / Q000276:immunology / Q000494:pharmacology; D020871:RNA Stability; D012333:RNA, Messenger / Q000737:chemistry*; D014158:Transcription, Genetic; D014529:Uridine / Q000031:analogs & derivatives / Q000737:chemistry
C103085:1-(3-C-ethynylribopentofuranosyl)uracil; D012333:RNA, Messenger; D016899:Interferon-beta; D011070:Poly I-C; D014529:Uridine