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Cryo-EM Studies of Pre-mRNA Splicing: From Sample Preparation to Model Visualization.
- Source :
-
Annual Review of Biophysics . May2018, Vol. 47, p175-199. 22p. - Publication Year :
- 2018
-
Abstract
- The removal of noncoding introns from pre-messenger RNA (pre-mRNA) is an essential step in eukaryotic gene expression and is catalyzed by a dynamic multi-megadalton ribonucleoprotein complex called the spliceosome. The spliceosome assembles on pre-mRNA substrates by the stepwise addition of small nuclear ribonucleoprotein particles and numerous protein factors. Extensive remodeling is required to form the RNA-based active site and to mediate the pre-mRNA branching and ligation reactions. In the past two years, cryo-electron microscopy (cryo-EM) structures of spliceosomes captured in different assembly and catalytic states have greatly advanced our understanding of its mechanism. This was made possible by long-standing efforts in the purification of spliceosome intermediates as well as recent developments in cryo-EM imaging and computational methodology. The resulting high-resolution densities allow for de novo model building in core regions of the complexes. In peripheral and less ordered regions, the combination of cross-linking, bioinformatics, biochemical, and genetic data is essential for accurate modeling. Here, we summarize these achievements and highlight the critical steps in obtaining near-atomic resolution structures of the spliceosome. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 1936122X
- Volume :
- 47
- Database :
- Academic Search Index
- Journal :
- Annual Review of Biophysics
- Publication Type :
- Academic Journal
- Accession number :
- 129752265
- Full Text :
- https://doi.org/10.1146/annurev-biophys-070317-033410