TY  - JOUR
A1  - Windhager, Lukas
A1  - Bonfert, Thomas
A1  - Burger, Kaspar
A1  - Ruzsics, Zsolt
A1  - Krebs, Stefan
A1  - Kaufmann, Stefanie
A1  - Malterer, Georg
A1  - L'Hernault, Anne
A1  - Schilhabel, Markus
A1  - Schreiber, Stefan
A1  - Rosenstiel, Philip
A1  - Zimmer, Ralf
A1  - Eick, Dirk
A1  - Friedel, Caroline C.
A1  - Dölken, Lars
T1  - Ultrashort and progressive 4sU-tagging reveals key characteristics of RNA processing at nucleotide resolution
Y1  - 2012/10/01 
JF  - Genome Research 
JO  - Genome Research 
SP  - 2031 
EP  - 2042 
DO  - 10.1101/gr.131847.111 
VL  - 22 
IS  - 10 
UR  - http://genome.cshlp.org/content/22/10/2031.abstract 
N2  - 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. 
ER  -