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

0351126 - ÚMG 2011 RIV US eng J - Journal Article
Huranová, Martina - Ivani, Ivan - Benda, Aleš - Poser, I. - Brody, Y. - Hof, Martin - Shav-Tal, Y. - Neugebauer, K.M. - Staněk, David
The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells.
Journal of Cell Biology. Roč. 191, č. 1 (2010), s. 75-86. ISSN 0021-9525. E-ISSN 1540-8140
R&D Projects: GA ČR GA204/07/0133; GA ČR GAP305/10/0424; GA AV ČR KAN200520801; GA MŠMT(CZ) LC06063
Institutional research plan: CEZ:AV0Z50520514; CEZ:AV0Z40400503
Keywords : FRAP * RNA splicing * spliceosome
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 9.921, year: 2010

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.
Permanent Link: http://hdl.handle.net/11104/0190946