Publication
Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
| dc.contributor.author | Martin, Robert M. | |
| dc.contributor.author | Rino, José | |
| dc.contributor.author | Carvalho, Célia | |
| dc.contributor.author | Kirchhausen, Tomas | |
| dc.contributor.author | Carmo-Fonseca, Maria | |
| dc.date.accessioned | 2021-12-03T15:47:59Z | |
| dc.date.available | 2021-12-03T15:47:59Z | |
| dc.date.issued | 2013 | |
| dc.description | © 2013 The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited. | pt_PT |
| dc.description.abstract | Removal of introns from pre-messenger RNAs (pre-mRNAs) via splicing provides a versatile means of genetic regulation that is often disrupted in human diseases. To decipher how splicing occurs in real time, we directly examined with single-molecule sensitivity the kinetics of intron excision from pre-mRNA in the nucleus of living human cells. By using two different RNA labeling methods, MS2 and λN, we show that β-globin introns are transcribed and excised in 20-30 s. Furthermore, we show that replacing the weak polypyrimidine (Py) tract in mouse immunoglobulin μ (IgM) pre-mRNA by a U-rich Py decreases the intron lifetime, thus providing direct evidence that splice-site strength influences splicing kinetics. We also found that RNA polymerase II transcribes at elongation rates ranging between 3 and 6 kb min(-1) and that transcription can be rate limiting for splicing. These results have important implications for a mechanistic understanding of cotranscriptional splicing regulation in the live-cell context. | pt_PT |
| dc.description.sponsorship | This work was supported by a grant from Fundação para a Ciência e Tecnologia, Portugal (PTDC/SAU-GMG/118180/2010), and the Harvard Medical School-Portugal Program in Translational Research and Information. R.M.M. was supported by a grant from Fundação para a Ciência e Tecnologia, Portugal (SFRH/BPD/66611/2009), and received an EMBO Short Term Fellowship (ASTF272.00-2011). J.R. holds a Ciência 2008 position from the Portuguese Ministry of Science and Technology. E.M. was supported in part by grants from the National Institutes of Health (NIH U54 AI057159) and the New England Regional Center of Excellence in Biodefense and Emerging Infectious Diseases. T.K. was supported in part by NIH grant GM 075252. | pt_PT |
| dc.description.version | info:eu-repo/semantics/publishedVersion | pt_PT |
| dc.identifier.citation | Cell reports, 4(6), 1144–1155 | pt_PT |
| dc.identifier.doi | 10.1016/j.celrep.2013.08.013 | pt_PT |
| dc.identifier.eissn | 2639-1856 | |
| dc.identifier.issn | 2211-1247 | |
| dc.identifier.uri | http://hdl.handle.net/10451/50271 | |
| dc.language.iso | eng | pt_PT |
| dc.peerreviewed | yes | pt_PT |
| dc.publisher | Elsevier | pt_PT |
| dc.relation | Splicing and stochasticity in programmed cell death decisions | |
| dc.relation | A SYNTHETIC BIOLOGY APPROACH TO STUDY SPLICING REGULATION | |
| dc.relation.publisherversion | https://www.sciencedirect.com/journal/cell-reports | pt_PT |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | pt_PT |
| dc.title | Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity | pt_PT |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.awardTitle | Splicing and stochasticity in programmed cell death decisions | |
| oaire.awardTitle | A SYNTHETIC BIOLOGY APPROACH TO STUDY SPLICING REGULATION | |
| oaire.awardURI | info:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FSAU-GMG%2F118180%2F2010/PT | |
| oaire.awardURI | info:eu-repo/grantAgreement/FCT//SFRH%2FBPD%2F66611%2F2009/PT | |
| oaire.citation.endPage | 1155 | pt_PT |
| oaire.citation.issue | 6 | pt_PT |
| oaire.citation.startPage | 1144 | pt_PT |
| oaire.citation.title | Cell Reports | pt_PT |
| oaire.citation.volume | 4 | pt_PT |
| oaire.fundingStream | 3599-PPCDT | |
| person.familyName | Martin | |
| person.familyName | Rino Henriques | |
| person.familyName | Carvalho | |
| person.familyName | Carmo-Fonseca | |
| person.givenName | Robert Manfred | |
| person.givenName | José Miguel | |
| person.givenName | Célia | |
| person.givenName | Maria | |
| person.identifier.ciencia-id | 7516-6C51-460B | |
| person.identifier.ciencia-id | A117-FBDB-A27B | |
| person.identifier.ciencia-id | DD18-667D-59FE | |
| person.identifier.ciencia-id | B31F-0435-0753 | |
| person.identifier.orcid | 0000-0001-5600-7525 | |
| person.identifier.orcid | 0000-0002-0637-4527 | |
| person.identifier.orcid | 0000-0002-5980-595X | |
| person.identifier.orcid | 0000-0002-3402-7143 | |
| person.identifier.scopus-author-id | 7007128195 | |
| project.funder.identifier | http://doi.org/10.13039/501100001871 | |
| project.funder.identifier | http://doi.org/10.13039/501100001871 | |
| project.funder.name | Fundação para a Ciência e a Tecnologia | |
| project.funder.name | Fundação para a Ciência e a Tecnologia | |
| rcaap.rights | openAccess | pt_PT |
| rcaap.type | article | pt_PT |
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