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PMID: 26829225 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cryo-EM structure of the yeast U4/U6.U5 tri-snRNP at 3.7 Å resolution.

Nature ·Vol. 530 ·No. 7590 ·2016-02-18 ·Pages 298-302

Nguyen THD, Galej WP, Bai XC, Oubridge C, Newman AJ, Scheres SHW, Nagai K

Abstract

U4/U6.U5 tri-snRNP represents a substantial part of the spliceosome before activation. A cryo-electron microscopy structure of Saccharomyces cerevisiae U4/U6.U5 tri-snRNP at 3.7 Å resolution led to an essentially complete atomic model comprising 30 proteins plus U4/U6 and U5 small nuclear RNAs (snRNAs). The structure reveals striking interweaving interactions of the protein and RNA components, including extended polypeptides penetrating into subunit interfaces. The invariant ACAGAGA sequence of U6 snRNA, which base-pairs with the 5'-splice site during catalytic activation, forms a hairpin stabilized by Dib1 and Prp8 while the adjacent nucleotides interact with the exon binding loop 1 of U5 snRNA. Snu114 harbours GTP, but its putative catalytic histidine is held away from the γ-phosphate by hydrogen bonding to a tyrosine in the amino-terminal domain of Prp8. Mutation of this histidine to alanine has no detectable effect on yeast growth. The structure provides important new insights into the spliceosome activation process leading to the formation of the catalytic centre.

MeSH Terms
Base Pairing Catalytic Domain Cryoelectron Microscopy DNA Helicases/metabolism Exons/genetics Guanosine Triphosphate/metabolism Hydrogen Bonding Models, Molecular Nucleic Acid Conformation RNA Splice Sites RNA, Small Nuclear/chemistry,genetics,metabolism Ribonucleoprotein, U4-U6 Small Nuclear/chemistry,metabolism Ribonucleoprotein, U5 Small Nuclear/chemistry,metabolism Ribonucleoproteins, Small Nuclear/chemistry,genetics,metabolism,ultrastructure Saccharomyces cerevisiae/chemistry,genetics,growth & development,ultrastructure Saccharomyces cerevisiae Proteins/chemistry,genetics,metabolism,ultrastructure Spliceosomes/metabolism
Chemicals
(U4-U6.U5)tri-snRNP-specific protein, S cerevisiae PRP8 protein, S cerevisiae RNA Splice Sites RNA, Small Nuclear Ribonucleoprotein, U4-U6 Small Nuclear Ribonucleoprotein, U5 Small Nuclear Ribonucleoproteins, Small Nuclear SNU114 protein, S cerevisiae Saccharomyces cerevisiae Proteins U5 small nuclear RNA Guanosine Triphosphate HCS1 protein, S cerevisiae DNA 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.
Oubridge Chris
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/nature16940
Published
2016-02-18
Epub
2016-00-01
Pages
298-302
Language
English
Region
England
NLM ID
0410462
PMCID
PMC4762201
Subset
IM
Grants
Medical Research Council · MC_U105184330 · United Kingdom
Medical Research Council · MC_UP_A025_1013 · United Kingdom
Databases
PDB
5GAM, 5GAN, 5GAO, 5GAP
External Links
PubMed source DOI: 10.1038/nature16940 PMC Full Text View journal details
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