کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1929024 | 1050441 | 2012 | 6 صفحه PDF | دانلود رایگان |
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.
► RNA metabolic labeling by EU provides accurate analysis of transcriptional activity.
► Transcriptional activity is maintained after the maximum accumulation of IFN-β mRNA.
► mRNA stability can be analyzed by EU labeling without interfering cell physiology.
► Transcriptional timings might determine the stability of interferon-beta mRNA.
► EU labeling is a useful tool to study gene expression regulation.
Journal: Biochemical and Biophysical Research Communications - Volume 428, Issue 1, 9 November 2012, Pages 44–49