Windhager, L.* ; Bonfert, T.* ; Burger, K. ; Ruzsics, Z.* ; Krebs, S.* ; Kaufmann, S.* ; Malterer, G.* ; L'hernault, A.* ; Schilhabel, M.* ; Schreiber, S.* ; Rosenstiel, P.* ; Zimmer, R.* ; Eick, D. ; Friedel, C.C.* ; Dölken, L.*
Ultrashort and progressive 4sU-tagging reveals key characteristics of RNA processing at nucleotide resolution.
Genome Res. 22, 2031-2042 (2012)
RNA synthesis and decay rates determine the steady-state levels of cellular RNAs. Metabolic tagging of newly transcribed RNA by 4-thiouridine (4sU) can reveal the relative contributions of RNA synthesis and decay rates. The kinetics of RNA processing, however, had so far remained unresolved. Here, we show that ultrashort 4sU-tagging not only provides snapshot pictures of eukaryotic gene expression but, when combined with progressive 4sU-tagging and RNA-seq, reveals global RNA processing kinetics at nucleotide resolution. Using this method, we identified classes of rapidly and slowly spliced/degraded introns. Interestingly, each class of splicing kinetics was characterized by a distinct association with intron length, gene length, and splice site strength. For a large group of introns, we also observed long lasting retention in the primary transcript, but efficient secondary splicing or degradation at later time points. Finally, we show that processing of most, but not all small nucleolar (sno)RNA-containing introns is remarkably inefficient with the majority of introns being spliced and degraded rather than processed into mature snoRNAs. In summary, our study yields unparalleled insights into the kinetics of RNA processing and provides the tools to study molecular mechanisms of RNA processing and their contribution to the regulation of gene expression.
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Publication type
Article: Journal article
Document type
Scientific Article
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Keywords
4sU-tagging; newly transcribed RNA; RNA-seq; RNA processing; 4-thiouridine; non-coding RNA; NEWLY SYNTHESIZED RNA; NONCODING RNAS; MICROARRAY ANALYSIS; MAMMALIAN-CELLS; POLYMERASE-II; MICRORNA; INTRON; IDENTIFICATION; TRANSCRIPTOME; ORGANIZATION
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Language
english
Publication Year
2012
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2012
ISSN (print) / ISBN
1088-9051
e-ISSN
1549-5469
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Volume: 22,
Issue: 10,
Pages: 2031-2042
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Cold Spring Harbor Laboratory Press
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Peer reviewed
POF-Topic(s)
30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
Research field(s)
Immune Response and Infection
PSP Element(s)
G-501490-001
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Erfassungsdatum
2012-11-01