A composite double-/single-stranded RNA-binding region in protein Prp3 supports tri-snRNP stability and splicing
- Freie Universität Berlin, Germany
- Max Planck Institute for Biophysical Chemistry, Germany
- International Institute of Molecular and Cell Biology, Poland
Abstract
Prp3 is an essential U4/U6 di-snRNP-associated protein whose functions and molecular mechanisms in pre-mRNA splicing are presently poorly understood. We show by structural and biochemical analyses that Prp3 contains a bipartite U4/U6 di-snRNA-binding region comprising an expanded ferredoxin-like fold, which recognizes a 3'-overhang of U6 snRNA, and a preceding peptide, which binds U4/U6 stem II. Phylogenetic analyses revealed that the single-stranded RNA-binding domain is exclusively found in Prp3 orthologs, thus qualifying as a spliceosome-specific RNA interaction module. The composite double-stranded/single-stranded RNA-binding region assembles cooperatively with Snu13 and Prp31 on U4/U6 di-snRNAs and inhibits Brr2-mediated U4/U6 di-snRNA unwinding in vitro. RNP-disrupting mutations in Prp3 lead to U4/U6•U5 tri-snRNP assembly and splicing defects in vivo. Our results reveal how Prp3 acts as an important bridge between U4/U6 and U5 in the tri-snRNP and comparison with a Prp24-U6 snRNA recycling complex suggests how Prp3 may be involved in U4/U6 re-assembly after splicing.
Article and author information
Author details
Reviewing Editor
- Timothy W Nilsen, Case Western Reserve University, United States
Version history
- Received: March 5, 2015
- Accepted: July 9, 2015
- Accepted Manuscript published: July 10, 2015 (version 1)
- Version of Record published: July 30, 2015 (version 2)
Copyright
© 2015, Liu et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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A composite double-/single-stranded RNA-binding region in protein Prp3 supports tri-snRNP stability and splicing
eLife 4:e07320.
https://doi.org/10.7554/eLife.07320
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