The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells

Precursor messenger RNA (pre-mRNA) splicing is catalyzed by the spliceosome, a large ribonucleoprotein (RNP) complex composed of five small nuclear RNP particles (snRNPs) and additional proteins. Using live cell imaging of GFP-tagged snRNP components expressed at endogenous levels, we examined how t...

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Detalles Bibliográficos
Autores principales: Huranová, Martina, Ivani, Ivan, Benda, Aleš, Poser, Ina, Brody, Yehuda, Hof, Martin, Shav-Tal, Yaron, Neugebauer, Karla M., Staněk, David
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2010
Materias:
Research Articles
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2953428/
https://www.ncbi.nlm.nih.gov/pubmed/20921136
http://dx.doi.org/10.1083/jcb.201004030
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author Huranová, Martina
Ivani, Ivan
Benda, Aleš
Poser, Ina
Brody, Yehuda
Hof, Martin
Shav-Tal, Yaron
Neugebauer, Karla M.
Staněk, David
author_facet Huranová, Martina
Ivani, Ivan
Benda, Aleš
Poser, Ina
Brody, Yehuda
Hof, Martin
Shav-Tal, Yaron
Neugebauer, Karla M.
Staněk, David
author_sort Huranová, Martina
collection PubMed
description Precursor messenger RNA (pre-mRNA) splicing is catalyzed by the spliceosome, a large ribonucleoprotein (RNP) complex composed of five small nuclear RNP particles (snRNPs) and additional proteins. Using live cell imaging of GFP-tagged snRNP components expressed at endogenous levels, we examined how the spliceosome assembles in vivo. A comprehensive analysis of snRNP dynamics in the cell nucleus enabled us to determine snRNP diffusion throughout the nucleoplasm as well as the interaction rates of individual snRNPs with pre-mRNA. Core components of the spliceosome, U2 and U5 snRNPs, associated with pre-mRNA for 15–30 s, indicating that splicing is accomplished within this time period. Additionally, binding of U1 and U4/U6 snRNPs with pre-mRNA occurred within seconds, indicating that the interaction of individual snRNPs with pre-mRNA is distinct. These results are consistent with the predictions of the step-wise model of spliceosome assembly and provide an estimate on the rate of splicing in human cells.
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spelling pubmed-29534282011-04-04 The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells Huranová, Martina Ivani, Ivan Benda, Aleš Poser, Ina Brody, Yehuda Hof, Martin Shav-Tal, Yaron Neugebauer, Karla M. Staněk, David J Cell Biol Research Articles Precursor messenger RNA (pre-mRNA) splicing is catalyzed by the spliceosome, a large ribonucleoprotein (RNP) complex composed of five small nuclear RNP particles (snRNPs) and additional proteins. Using live cell imaging of GFP-tagged snRNP components expressed at endogenous levels, we examined how the spliceosome assembles in vivo. A comprehensive analysis of snRNP dynamics in the cell nucleus enabled us to determine snRNP diffusion throughout the nucleoplasm as well as the interaction rates of individual snRNPs with pre-mRNA. Core components of the spliceosome, U2 and U5 snRNPs, associated with pre-mRNA for 15–30 s, indicating that splicing is accomplished within this time period. Additionally, binding of U1 and U4/U6 snRNPs with pre-mRNA occurred within seconds, indicating that the interaction of individual snRNPs with pre-mRNA is distinct. These results are consistent with the predictions of the step-wise model of spliceosome assembly and provide an estimate on the rate of splicing in human cells. The Rockefeller University Press 2010-10-04 /pmc/articles/PMC2953428/ /pubmed/20921136 http://dx.doi.org/10.1083/jcb.201004030 Text en © 2010 Huranová et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Research Articles
Huranová, Martina
Ivani, Ivan
Benda, Aleš
Poser, Ina
Brody, Yehuda
Hof, Martin
Shav-Tal, Yaron
Neugebauer, Karla M.
Staněk, David
The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells
title The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells
title_full The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells
title_fullStr The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells
title_full_unstemmed The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells
title_short The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells
title_sort differential interaction of snrnps with pre-mrna reveals splicing kinetics in living cells
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2953428/
https://www.ncbi.nlm.nih.gov/pubmed/20921136
http://dx.doi.org/10.1083/jcb.201004030
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