Kinetic competition during the transcription cycle results in stochastic RNA processing
Synthesis of mRNA in eukaryotes involves the coordinated action of many enzymatic processes, including initiation, elongation, splicing, and cleavage. Kinetic competition between these processes has been proposed to determine RNA fate, yet such coupling has never been observed in vivo on single tran...
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| Format: | Online |
| Language: | English |
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eLife Sciences Publications, Ltd
2014
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4210818/ |
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pubmed-4210818 |
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pubmed-42108182014-11-21 Kinetic competition during the transcription cycle results in stochastic RNA processing Coulon, Antoine Ferguson, Matthew L de Turris, Valeria Palangat, Murali Chow, Carson C Larson, Daniel R Biophysics and Structural Biology Synthesis of mRNA in eukaryotes involves the coordinated action of many enzymatic processes, including initiation, elongation, splicing, and cleavage. Kinetic competition between these processes has been proposed to determine RNA fate, yet such coupling has never been observed in vivo on single transcripts. In this study, we use dual-color single-molecule RNA imaging in living human cells to construct a complete kinetic profile of transcription and splicing of the β-globin gene. We find that kinetic competition results in multiple competing pathways for pre-mRNA splicing. Splicing of the terminal intron occurs stochastically both before and after transcript release, indicating there is not a strict quality control checkpoint. The majority of pre-mRNAs are spliced after release, while diffusing away from the site of transcription. A single missense point mutation (S34F) in the essential splicing factor U2AF1 which occurs in human cancers perturbs this kinetic balance and defers splicing to occur entirely post-release. eLife Sciences Publications, Ltd 2014-10-01 /pmc/articles/PMC4210818/ /pubmed/25271374 http://dx.doi.org/10.7554/eLife.03939 Text en http://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication (http://creativecommons.org/publicdomain/zero/1.0) . |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Coulon, Antoine Ferguson, Matthew L de Turris, Valeria Palangat, Murali Chow, Carson C Larson, Daniel R |
| spellingShingle |
Coulon, Antoine Ferguson, Matthew L de Turris, Valeria Palangat, Murali Chow, Carson C Larson, Daniel R Kinetic competition during the transcription cycle results in stochastic RNA processing |
| author_facet |
Coulon, Antoine Ferguson, Matthew L de Turris, Valeria Palangat, Murali Chow, Carson C Larson, Daniel R |
| author_sort |
Coulon, Antoine |
| title |
Kinetic competition during the transcription cycle results in stochastic RNA processing |
| title_short |
Kinetic competition during the transcription cycle results in stochastic RNA processing |
| title_full |
Kinetic competition during the transcription cycle results in stochastic RNA processing |
| title_fullStr |
Kinetic competition during the transcription cycle results in stochastic RNA processing |
| title_full_unstemmed |
Kinetic competition during the transcription cycle results in stochastic RNA processing |
| title_sort |
kinetic competition during the transcription cycle results in stochastic rna processing |
| description |
Synthesis of mRNA in eukaryotes involves the coordinated action of many enzymatic processes, including initiation, elongation, splicing, and cleavage. Kinetic competition between these processes has been proposed to determine RNA fate, yet such coupling has never been observed in vivo on single transcripts. In this study, we use dual-color single-molecule RNA imaging in living human cells to construct a complete kinetic profile of transcription and splicing of the β-globin gene. We find that kinetic competition results in multiple competing pathways for pre-mRNA splicing. Splicing of the terminal intron occurs stochastically both before and after transcript release, indicating there is not a strict quality control checkpoint. The majority of pre-mRNAs are spliced after release, while diffusing away from the site of transcription. A single missense point mutation (S34F) in the essential splicing factor U2AF1 which occurs in human cancers perturbs this kinetic balance and defers splicing to occur entirely post-release. |
| publisher |
eLife Sciences Publications, Ltd |
| publishDate |
2014 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4210818/ |
| _version_ |
1613149243892039680 |