Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity

Detalhes bibliográficos
Autor(a) principal: Martin, Robert M.
Data de Publicação: 2013
Outros Autores: Rino, José, Carvalho, Célia, Kirchhausen, Tomas, Carmo-Fonseca, Maria
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Texto Completo: http://hdl.handle.net/10451/50271
Resumo: © 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.
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spelling Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity© 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.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.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.ElsevierRepositório da Universidade de LisboaMartin, Robert M.Rino, JoséCarvalho, CéliaKirchhausen, TomasCarmo-Fonseca, Maria2021-12-03T15:47:59Z20132013-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10451/50271engCell reports, 4(6), 1144–11552211-124710.1016/j.celrep.2013.08.0132639-1856info:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2025-03-17T14:38:37Zoai:repositorio.ulisboa.pt:10451/50271Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-29T03:19:30.523088Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
title Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
spellingShingle Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
Martin, Robert M.
title_short Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
title_full Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
title_fullStr Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
title_full_unstemmed Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
title_sort Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
author Martin, Robert M.
author_facet Martin, Robert M.
Rino, José
Carvalho, Célia
Kirchhausen, Tomas
Carmo-Fonseca, Maria
author_role author
author2 Rino, José
Carvalho, Célia
Kirchhausen, Tomas
Carmo-Fonseca, Maria
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Martin, Robert M.
Rino, José
Carvalho, Célia
Kirchhausen, Tomas
Carmo-Fonseca, Maria
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.
publishDate 2013
dc.date.none.fl_str_mv 2013
2013-01-01T00:00:00Z
2021-12-03T15:47:59Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10451/50271
url http://hdl.handle.net/10451/50271
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Cell reports, 4(6), 1144–1155
2211-1247
10.1016/j.celrep.2013.08.013
2639-1856
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron:RCAAP
instname_str FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron_str RCAAP
institution RCAAP
reponame_str Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
collection Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
repository.name.fl_str_mv Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
repository.mail.fl_str_mv info@rcaap.pt
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