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The composition of the transcriptome is determined by a balance between mRNA synthesis and degradation. An important route for mRNA degradation produces uncapped mRNAs, and this decay process can be initiated by decapping enzymes, endonucleases, and small RNAs. Although uncapped mRNAs are an important intermediate for mRNA decay, their identity and abundance have never been studied on a large scale until recently. Here, we present an experimental method for transcriptome-wide profiling of uncapped mRNAs that can be used in any eukaryotic system. We applied the method to study the prevalence of uncapped transcripts during the early stages of Arabidopsis thaliana flower development. Uncapped transcripts were identified for the majority of expressed genes, although at different levels. By comparing uncapped RNA levels with steady state overall transcript levels, our study provides evidence for widespread mRNA degradation control in numerous biological processes involving genes of varied molecular functions, implying that uncapped mRNA levels are dynamically regulated. Sequence analyses identified structural features of transcripts and cis-elements that were associated with different levels of uncapping. These transcriptome-wide profiles of uncapped mRNAs will aid in illuminating new regulatory mechanisms of eukaryotic transcriptional networks.
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http://dx.doi.org/10.1105/tpc.108.062786 | DOI Listing |
Mol Ther Nucleic Acids
June 2025
Process and Analytical Sciences, BioPharmaceuticals Development, BioPharmaceuticals R&D, AstraZeneca, Cambridge CB2 0AA, UK.
The 5'Cap is a modified nucleotide structure added to the 5'end of mRNA, which plays a critical role in stability and translation of the molecule. Accurate characterization of 5'Cap impurities is crucial during development of mRNA therapeutics. Here, we utilized orthogonal liquid chromatography-mass spectrometry (LC-MS) workflows to characterize Cap-1-modified mRNA.
View Article and Find Full Text PDFBiochem Biophys Res Commun
June 2025
College of Biological Science and Medical Engineering, DongHua University Shanghai 201620, China. Electronic address:
Understanding and meticulously controlling mRNA capping efficiency is an indispensable quality attribute in developing mRNA therapeutics. A rigorously defined capping process is critical for ensuring the consistency and reproducibility of mRNA drugs, forming the cornerstone of compliance with stringent regulatory standards. This investigation introduces an innovative methodology for assessing mRNA's capping efficiency through applying the Pistol ribozyme and liquid chromatography-mass spectrometry (LC-MS).
View Article and Find Full Text PDFBiochim Biophys Acta Gene Regul Mech
June 2025
Institute of Health Sciences, Presidency University, Plot No. DG/02/02, Premises No. 14-0358, Action Area 1D, Kolkata 700156, India. Electronic address:
In response to hypoxia, hypoxia-inducible factors (HIFs) control the transcriptomic output to mitigate the hypoxic stress. Long noncoding RNAs (lncRNA) are found to be very crucial in regulating hypoxia. Like mRNAs, lncRNAs are protected by 5' caps that are added by mRNA capping enzyme (CE) in the nucleus.
View Article and Find Full Text PDFAnn Med
December 2025
College of Bioengineering, National ''111'' Center for Cellular Regulation and Molecular Pharmaceutics, Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Hubei University of Technology, Wuhan, China.
J Biol Chem
November 2024
Department of Chemistry, Hunter College, City University of New York, New York, New York, USA; PhD. Program in Biochemistry, The Graduate Center of the City University of New York, New York, New York, USA; PhD. Program in Chemistry, The Graduate Center of the City University of New York, New York, N
Protein translation is globally downregulated under stress conditions. Many proteins that are synthesized under stress conditions use a cap-independent translation initiation pathway. A subset of cellular mRNAs that encode for these proteins contain stable secondary structures within their 5'UTR, and initiate cap-independent translation using elements called cap-independent translation enhancers or internal ribosome entry sites within their 5'UTRs.
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