Abstract
Precursor messenger RNA (pre-mRNA) splicing is catalyzed by the spliceosome, a large ribonucleo-protein (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.
| Original language | English |
|---|---|
| Pages (from-to) | 75-86 |
| Number of pages | 12 |
| Journal | Journal of Cell Biology |
| Volume | 191 |
| Issue number | 1 |
| DOIs | |
| State | Published - 4 Oct 2010 |
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