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Structure paper

TitleCryo-EM structure of the spliceosome immediately after branching.
Journal, issue, pagesNature, Vol. 537, Issue 7619, Page 197-201, Year 2016
Publish dateSep 8, 2016
AuthorsWojciech P Galej / Max E Wilkinson / Sebastian M Fica / Chris Oubridge / Andrew J Newman / Kiyoshi Nagai /
PubMed AbstractPrecursor mRNA (pre-mRNA) splicing proceeds by two consecutive transesterification reactions via a lariat-intron intermediate. Here we present the 3.8 Å cryo-electron microscopy structure of the ...Precursor mRNA (pre-mRNA) splicing proceeds by two consecutive transesterification reactions via a lariat-intron intermediate. Here we present the 3.8 Å cryo-electron microscopy structure of the spliceosome immediately after lariat formation. The 5'-splice site is cleaved but remains close to the catalytic Mg site in the U2/U6 small nuclear RNA (snRNA) triplex, and the 5'-phosphate of the intron nucleotide G(+1) is linked to the branch adenosine 2'OH. The 5'-exon is held between the Prp8 amino-terminal and linker domains, and base-pairs with U5 snRNA loop 1. Non-Watson-Crick interactions between the branch helix and 5'-splice site dock the branch adenosine into the active site, while intron nucleotides +3 to +6 base-pair with the U6 snRNA ACAGAGA sequence. Isy1 and the step-one factors Yju2 and Cwc25 stabilize docking of the branch helix. The intron downstream of the branch site emerges between the Prp8 reverse transcriptase and linker domains and extends towards the Prp16 helicase, suggesting a plausible mechanism of remodelling before exon ligation.
External linksNature / PubMed:27459055 / PubMed Central
MethodsEM (single particle)
Resolution3.8 - 10.0 Å
Structure data

4055

5lj3

EMDB-4055: Overall map of the yeast spliceosome immediately after branching
PDB-5lj3: Structure of the core of the yeast spliceosome immediately after branching
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-4056:
Structure of the yeast spliceosome immediately after branching. 3D class containing Prp19 module.
Method: EM (single particle) / Resolution: 5.1 Å

4057

5lj5

EMDB-4057: Structure of the yeast spliceosome immediately after branching. 3D class containing helicase module.
PDB-5lj5: Overall structure of the yeast spliceosome immediately after branching.
Method: EM (single particle) / Resolution: 10.0 Å

EMDB-4058:
Structure of the yeast spliceosome immediately after branching. Core without WD40 domain.
Method: EM (single particle) / Resolution: 4.1 Å

EMDB-4059:
Structure of the yeast spliceosome immediately after branching. Core with WD40 domain.
Method: EM (single particle) / Resolution: 3.9 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM / Guanosine triphosphate

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • Saccharomyces cerevisiae
  • Baker's yeast (brewer's yeast)
KeywordsSPLICING / spliceosome / snRNP / pre-mRNA splicing / trans-esterification / lariat intermediate / complex C

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