Dynamic imaging of nascent RNA reveals general principles of transcription dynamics and stochastic splice site selection.
RNA
fluorescence
heterogeneity
imaging
single molecule
spliceosome
splicing
stochastic
transcription
Journal
Cell
ISSN: 1097-4172
Titre abrégé: Cell
Pays: United States
ID NLM: 0413066
Informations de publication
Date de publication:
27 05 2021
27 05 2021
Historique:
received:
16
09
2019
revised:
12
11
2020
accepted:
08
04
2021
pubmed:
13
5
2021
medline:
22
12
2021
entrez:
12
5
2021
Statut:
ppublish
Résumé
The activities of RNA polymerase and the spliceosome are responsible for the heterogeneity in the abundance and isoform composition of mRNA in human cells. However, the dynamics of these megadalton enzymatic complexes working in concert on endogenous genes have not been described. Here, we establish a quasi-genome-scale platform for observing synthesis and processing kinetics of single nascent RNA molecules in real time. We find that all observed genes show transcriptional bursting. We also observe large kinetic variation in intron removal for single introns in single cells, which is inconsistent with deterministic splice site selection. Transcriptome-wide footprinting of the U2AF complex, nascent RNA profiling, long-read sequencing, and lariat sequencing further reveal widespread stochastic recursive splicing within introns. We propose and validate a unified theoretical model to explain the general features of transcription and pervasive stochastic splice site selection.
Identifiants
pubmed: 33979654
pii: S0092-8674(21)00491-8
doi: 10.1016/j.cell.2021.04.012
pmc: PMC8183334
mid: NIHMS1700536
pii:
doi:
Substances chimiques
RNA Precursors
0
RNA Splice Sites
0
RNA, Messenger
0
Types de publication
Journal Article
Research Support, N.I.H., Intramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2878-2895.e20Subventions
Organisme : Intramural NIH HHS
ID : Z99 CA999999
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIA BC011383
Pays : United States
Commentaires et corrections
Type : CommentIn
Informations de copyright
Published by Elsevier Inc.
Déclaration de conflit d'intérêts
Declaration of interests The authors declare no competing interests.
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