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The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells
M. Huranová, I. Ivani, A. Benda, I. Poser, Y. Brody, M. Hof, Y. Shav-Tal, KM. Neugebauer, D. Stanek,
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
NLK
Free Medical Journals
from 1962 to 6 months ago
Freely Accessible Science Journals
from 1962 to 6 months ago
Europe PubMed Central
from 1962 to 6 months ago
Open Access Digital Library
from 1955-01-25
Open Access Digital Library
from 1959-01-01
Open Access Digital Library
from 1962-01-01
Medline Complete (EBSCOhost)
from 2005-03-28 to 2011-09-19
PubMed
20921136
DOI
10.1083/jcb.201004030
Knihovny.cz E-resources
- MeSH
- Cell Nucleus metabolism MeSH
- Cell Line MeSH
- Spectrometry, Fluorescence MeSH
- Fluorescence Recovery After Photobleaching MeSH
- HeLa Cells MeSH
- Kinetics MeSH
- Humans MeSH
- RNA, Messenger metabolism MeSH
- RNA Precursors metabolism MeSH
- Ribonucleoproteins, Small Nuclear metabolism physiology MeSH
- RNA Splicing physiology MeSH
- Spliceosomes metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
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.
References provided by Crossref.org
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