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PMID: 26106855 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The architecture of the spliceosomal U4/U6.U5 tri-snRNP.

Nature ·Vol. 523 ·No. 7558 ·2015-07-02 ·Pages 47-52

Nguyen TH, Galej WP, Bai XC, Savva CG, Newman AJ, Scheres SH, Nagai K

Abstract

U4/U6.U5 tri-snRNP is a 1.5-megadalton pre-assembled spliceosomal complex comprising U5 small nuclear RNA (snRNA), extensively base-paired U4/U6 snRNAs and more than 30 proteins, including the key components Prp8, Brr2 and Snu114. The tri-snRNP combines with a precursor messenger RNA substrate bound to U1 and U2 small nuclear ribonucleoprotein particles (snRNPs), and transforms into a catalytically active spliceosome after extensive compositional and conformational changes triggered by unwinding of the U4 and U6 (U4/U6) snRNAs. Here we use cryo-electron microscopy single-particle reconstruction of Saccharomyces cerevisiae tri-snRNP at 5.9 Å resolution to reveal the essentially complete organization of its RNA and protein components. The single-stranded region of U4 snRNA between its 3' stem-loop and the U4/U6 snRNA stem I is loaded into the Brr2 helicase active site ready for unwinding. Snu114 and the amino-terminal domain of Prp8 position U5 snRNA to insert its loop I, which aligns the exons for splicing, into the Prp8 active site cavity. The structure provides crucial insights into the activation process and the active site of the spliceosome.

MeSH Terms
Binding Sites Cryoelectron Microscopy Models, Molecular Protein Structure, Quaternary RNA Helicases/chemistry,metabolism Ribonucleoprotein, U4-U6 Small Nuclear/chemistry,metabolism Ribonucleoprotein, U5 Small Nuclear/chemistry,metabolism Saccharomyces cerevisiae/chemistry,metabolism Saccharomyces cerevisiae Proteins/chemistry,metabolism Spliceosomes/chemistry,physiology
Chemicals
PRP8 protein, S cerevisiae Ribonucleoprotein, U4-U6 Small Nuclear Ribonucleoprotein, U5 Small Nuclear SNU114 protein, S cerevisiae Saccharomyces cerevisiae Proteins BRR2 protein, S cerevisiae RNA Helicases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Nguyen Thi Hoang Duong
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
Galej Wojciech P
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
Bai Xiao-chen
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
Savva Christos G
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
Newman Andrew J
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
Scheres Sjors H W
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
Nagai Kiyoshi
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
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Article Info
Journal
Nature
Abbr.
Nature
ISSN
1476-4687
DOI
10.1038/nature14548
Published
2015-07-02
Epub
2015-00-24
Pages
47-52
Language
English
Region
England
NLM ID
0410462
PMCID
PMC4536768
Subset
IM
Grants
Medical Research Council · MC_U105184330 · United Kingdom
Medical Research Council · MC_UP_A025_1013 · United Kingdom
External Links
PubMed source DOI: 10.1038/nature14548 PMC Full Text View journal details
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