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TitleThe architecture of the spliceosomal U4/U6.U5 tri-snRNP.
Journal, issue, pagesNature, Vol. 523, Issue 7558, Page 47-52, Year 2015
Publish dateJul 2, 2015
AuthorsThi Hoang Duong Nguyen / Wojciech P Galej / Xiao-chen Bai / Christos G Savva / Andrew J Newman / Sjors H W Scheres / Kiyoshi Nagai /
PubMed AbstractU4/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 ...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.
External linksNature / PubMed:26106855 / PubMed Central
MethodsEM (single particle)
Resolution5.9 Å
Structure data

EMDB-2966:
EM structure of the yeast spliceosomal U4/U6.U5 tri-snRNP
Method: EM (single particle) / Resolution: 5.9 Å

Source
  • Saccharomyces cerevisiae (baker's yeast)
KeywordsSpliceosome / tri-snRNP / BRR2 protein, S cerevisiae / Binding Sites / Cryoelectron Microscopy / Models, Molecular / PRP8 protein, S cerevisiae / Protein Structure, Quaternary / RNA Helicases / Ribonucleoprotein, U4-U6 Small Nuclear / Ribonucleoprotein, U5 Small Nuclear / SNU114 protein, S cerevisiae / Saccharomyces cerevisiae / Saccharomyces cerevisiae Proteins / Spliceosomes

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