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Structure paper

TitleStructures of the Catalytically Activated Yeast Spliceosome Reveal the Mechanism of Branching.
Journal, issue, pagesCell, Vol. 177, Issue 2, Page 339-351.e13, Year 2019
Publish dateApr 4, 2019
AuthorsRuixue Wan / Rui Bai / Chuangye Yan / Jianlin Lei / Yigong Shi /
PubMed AbstractPre-mRNA splicing is executed by the spliceosome. Structural characterization of the catalytically activated complex (B) is pivotal for understanding the branching reaction. In this study, we ...Pre-mRNA splicing is executed by the spliceosome. Structural characterization of the catalytically activated complex (B) is pivotal for understanding the branching reaction. In this study, we assembled the B complexes on two different pre-mRNAs from Saccharomyces cerevisiae and determined the cryo-EM structures of four distinct B complexes at overall resolutions of 2.9-3.8 Å. The duplex between U2 small nuclear RNA (snRNA) and the branch point sequence (BPS) is discretely away from the 5'-splice site (5'SS) in the three B complexes that are devoid of the step I splicing factors Yju2 and Cwc25. Recruitment of Yju2 into the active site brings the U2/BPS duplex into the vicinity of 5'SS, with the BPS nucleophile positioned 4 Å away from the catalytic metal M2. This analysis reveals the functional mechanism of Yju2 and Cwc25 in branching. These structures on different pre-mRNAs reveal substrate-specific conformations of the spliceosome in a major functional state.
External linksCell / PubMed:30879786
MethodsEM (single particle)
Resolution2.9 - 3.86 Å
Structure data

EMDB-0684:
Cryo-EM structure of the catalytic activated yeast spliceosome (B* complex) assembled on ACT1 pre-mRNA at 2.9 angstrom resolution
Method: EM (single particle) / Resolution: 2.9 Å

EMDB-0685:
Cryo-EM structure of the catalytic activated yeast spliceosome (B* complex) assembled on UBC4 pre-mRNA at 3.2 angstrom resolution
Method: EM (single particle) / Resolution: 3.2 Å

0686

6j6g

EMDB-0686, PDB-6j6g:
Cryo-EM structure of the yeast B*-a2 complex at an average resolution of 3.2 angstrom
Method: EM (single particle) / Resolution: 3.2 Å

0687

6j6h

EMDB-0687, PDB-6j6h:
Cryo-EM structure of the yeast B*-a1 complex at an average resolution of 3.6 angstrom
Method: EM (single particle) / Resolution: 3.6 Å

0691

6j6n

EMDB-0691, PDB-6j6n:
Cryo-EM structure of the yeast B*-b1 complex at an average resolution of 3.86 angstrom
Method: EM (single particle) / Resolution: 3.86 Å

0692

6j6q

EMDB-0692, PDB-6j6q:
Cryo-EM structure of the yeast B*-b2 complex at an average resolution of 3.7 angstrom
Method: EM (single particle) / Resolution: 3.7 Å

Chemicals

ChemComp-IHP:
INOSITOL HEXAKISPHOSPHATE

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

ChemComp-MG:
Unknown entry

ChemComp-ZN:
Unknown entry

Source
  • saccharomyces cerevisiae s288c (yeast)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • Baker's yeast (brewer's yeast)
KeywordsSPLICING / spliceosme / B* complex / branching / snRNP / U snRNA / spliceosome

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