Transcriptome-wide Interrogation of the Functional Intronome by Spliceosome Profiling.
Cell
; 173(4): 1031-1044.e13, 2018 05 03.
Article
en En
| MEDLINE
| ID: mdl-29727662
ABSTRACT
Full understanding of eukaryotic transcriptomes and how they respond to different conditions requires deep knowledge of all sites of intron excision. Although RNA sequencing (RNA-seq) provides much of this information, the low abundance of many spliced transcripts (often due to their rapid cytoplasmic decay) limits the ability of RNA-seq alone to reveal the full repertoire of spliced species. Here, we present "spliceosome profiling," a strategy based on deep sequencing of RNAs co-purifying with late-stage spliceosomes. Spliceosome profiling allows for unambiguous mapping of intron ends to single-nucleotide resolution and branchpoint identification at unprecedented depths. Our data reveal hundreds of new introns in S. pombe and numerous others that were previously misannotated. By providing a means to directly interrogate sites of spliceosome assembly and catalysis genome-wide, spliceosome profiling promises to transform our understanding of RNA processing in the nucleus, much as ribosome profiling has transformed our understanding mRNA translation in the cytoplasm.
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1
Banco de datos:
MEDLINE
Asunto principal:
Schizosaccharomyces
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Empalmosomas
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Transcriptoma
Idioma:
En
Año:
2018
Tipo del documento:
Article