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  Chemical Pull-Down Reveals Dynamic Pseudouridylation of the Mammalian Transcriptome

 
       Center of Life Sciences' research team lead by Dr. Chengqi Yi published their work entitled "Chemical Pull-Down Reveals Dynamic Pseudouridylation of the Mammalian Transcriptome " in Nature Chemical Biology on June 15th, 2015. In this study, the team reported CeU-Seq—a selective chemical labelling and pull-down method—to identify the transcriptome-wide, base-resolution pseudouridine (Y) RNA modification profiles within human cells and mouse tissues.
       More than 100 different types of post-transcriptional modifications to RNA molecules have been characterized so far. Among them, pseudouridine is overall the most abundant. The function and mechanism of pseudouridine in non-coding RNAs (rRNA, tRNA, snRNA and etc.) have been extensively studied; yet our knowledge regarding to the prevalence, mechanism and function of modifications in mRNA and long non-coding RNA (lncRNA) is limited. A major challenge is the lack of a selective and sensitive sequencing method for the transcriptome-wide identification of this post-transcriptional RNA modification.
       In this study, the team performed quantitative mass spectrometry analysis and show that pseudouridine is much more prevalent ( Y/U ratio: ~0.2% to 0.6%) in mammalian messenger RNA than previously believed. They developed CeU-Seq—a selective chemical labelling and pull-down method—to identify 2,084 Y sites within 1,929 human transcripts. They show that hPUS1, a known Y synthase, acts on human mRNA; upon stress, CeU-Seq demonstrates inducible and stress-specific mRNA pseudouridylation. Applications of CeU-Seq to mouse transcriptome reveal conserved and tissue-specific pseudouridylation. Collectively, these approaches allow comprehensive analysis of transcriptome-wide pseudouridylation and provide tools for functional studies of Y-mediated epigenetic regulation.
       The study was supported by the Ministry of Science and Technology, the National Natural Science Foundation of China, and Peking-Tsinghua Center for Life Sciences.
 
 
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