【摘 要】
：
Pseudouridine is the most abundant post-transcriptional RNA modification.Although pseudouridine is well established in non-coding RNAs, little is known about its prevalence and function in mRNA.To det
【机 构】
：
State Key Laboratory of Protein and Plant Gene Research,School of Life Sciences,and Peking-Tsinghua
【出 处】
：
2015 Genomics Frontiers Symposium(2015基因组学前沿研讨会)
论文部分内容阅读
Pseudouridine is the most abundant post-transcriptional RNA modification.Although pseudouridine is well established in non-coding RNAs, little is known about its prevalence and function in mRNA.To determine the pseudouridine content in mRNA, we performed quantitative mass spectrometry analysis in mammalian mRNAs using stringent mRNA purification procedures and found that pseudouridine is much more prevalent than previously believed.The ψ/U ratio ranges from 0.2% to 0.6%, which is comparable to the ratio of m6A in mRNA, the most abundant internal mRNA modification.To study the pseudouridine in the transcriptome, we developed N3-CMC-enriched pseudouridine sequencing (CeU-Seq), which is enabled via a chemically-synthesized CMC derivative that pre-enriches the ψ-containing RNA through biotin pulldown.Indeed, CeU-Seq allowed us to identify 2084 ψ sites in the transcriptome of HEK293T cells, and also 1741 and 1543 ψ sites in mouse brain and liver, respectively.Combining CeU-Seq and quantitative MS analysis,we showed that several pseudouridine synthases could act on mRNA and identified 77 PUS 1-dependent ψ sites.Furthermore, via CeU-Seq we identified several hundreds of stress-inducible ψ sites under heat shock or hydrogen peroxide conditions, respectively.Interestingly, the inducible ψ sites displayed strong stimulus-specific patterns and were essentially non-overlapping, indicating that mRNA pseudouridylation is dynamically regulated.Work is ongoing to map the ψ profiles of embryonic stem cells and model animals in different developmental stages.Functional roles of pseudouridine in different biological processes are also under investigation.
其他文献
Fibroblast growth factors (FGFs) make up a large family of polypeptide growth factors that are found in organisms ranging from nematodes to humans.In vertebrates, a number of FGFs have been shown to p
Chromosome conformation capture (3C) technology and its derivations, e.g., Hi-C, have made surveys of whole genome 3D structure possible, however, due to inherent limitations of the technology, it rem
RNA splicing is prevalent in mammalian transcriptome, and alternative splicing on pre-mRNAs is believed to be one of the major sources for protein divergence.However, it remains largely uncharacterize
Our previous study revealed that PDCD2 plays a role in erythroid differentiation (unpublished data).To get a further view, RNA-seq analyses of control and PDCD2-KD TF-1 cells were conducted, and 574 d
Transcriptome N6-methyladenine (m6A) methylation affects the regulation of various biological processes such as RNA stability and mRNA translation.Altered transcriptome methylation is known to contrib
Coral reefs represent one of the richest ocean micro-scale environments that include diverse collection of both basic and advanced species.Analysis of the microbial communities that live with coral re
Gliomas are one of the most common types of brain tumors.Numerous efforts have been contributed to dissect the mechanisms of glioma genesis and identify biomarkers for diagnosis and treatment.To facil
Cell sorting based method is widely used to screen clusters of cells during a complete biological process.We applied this technique to obtain populations of human erythroblasts at distinct development
Long non-coding RNAs (lncRNAs) are emerging as new regulators in the cancer paradigm,demonstrating potential roles in both oncogenic and tumor suppressive pathways.These novel genes are frequently abe
In eukaryotes, aberrant expression of transposable elements (TEs) is detrimental to the host genome.Piwi-interacting RNAs of~23 to 30 nucleotides (nt) bound to PIWI-clade Argonaute proteins silence tr