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The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells
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SYSNO ASEP 0351126 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells Author(s) Huranová, Martina (UMG-J) ORCID
Ivani, Ivan (UMG-J)
Benda, Aleš (UFCH-W) RID, ORCID
Poser, I. (DE)
Brody, Y. (IL)
Hof, Martin (UFCH-W) RID, ORCID
Shav-Tal, Y. (IL)
Neugebauer, K.M. (DE)
Staněk, David (UMG-J) RIDSource Title Journal of Cell Biology. - : Rockefeller University Press - ISSN 0021-9525
Roč. 191, č. 1 (2010), s. 75-86Number of pages 12 s. Language eng - English Country US - United States Keywords FRAP ; RNA splicing ; spliceosome Subject RIV EB - Genetics ; Molecular Biology R&D Projects GA204/07/0133 GA ČR - Czech Science Foundation (CSF) GAP305/10/0424 GA ČR - Czech Science Foundation (CSF) KAN200520801 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) LC06063 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) CEZ AV0Z50520514 - UMG-J (2005-2011) AV0Z40400503 - UFCH-W (2005-2011) UT WOS 000282648500009 DOI 10.1083/jcb.201004030 Annotation 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. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2011
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