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TitleStructural Insights into the Roles of Metazoan-Specific Splicing Factors in the Human Step 1 Spliceosome.
Journal, issue, pagesMol Cell, Vol. 80, Issue 1, Page 127-139.e6, Year 2020
Publish dateOct 1, 2020
AuthorsKarl Bertram / Leyla El Ayoubi / Olexandr Dybkov / Dmitry E Agafonov / Cindy L Will / Klaus Hartmuth / Henning Urlaub / Berthold Kastner / Holger Stark / Reinhard Lührmann /
PubMed AbstractHuman spliceosomes contain numerous proteins absent in yeast, whose functions remain largely unknown. Here we report a 3D cryo-EM structure of the human spliceosomal C complex at 3.4 Å core ...Human spliceosomes contain numerous proteins absent in yeast, whose functions remain largely unknown. Here we report a 3D cryo-EM structure of the human spliceosomal C complex at 3.4 Å core resolution and 4.5-5.7 Å at its periphery, and aided by protein crosslinking we determine its molecular architecture. Our structure provides additional insights into the spliceosome's architecture between the catalytic steps of splicing, and how proteins aid formation of the spliceosome's catalytically active RNP (ribonucleoprotein) conformation. It reveals the spatial organization of the metazoan-specific proteins PPWD1, WDR70, FRG1, and CIR1 in human C complexes, indicating they stabilize functionally important protein domains and RNA structures rearranged/repositioned during the B to C transition. Structural comparisons with human B, C, and P complexes reveal an intricate cascade of RNP rearrangements during splicing catalysis, with intermediate RNP conformations not found in yeast, and additionally elucidate the structural basis for the sequential recruitment of metazoan-specific spliceosomal proteins.
External linksMol Cell / PubMed:33007253
MethodsEM (single particle)
Resolution3.4 - 5.7 Å
Structure data

11569

6zym

EMDB-11569, PDB-6zym:
Human C Complex Spliceosome - High-resolution CORE
Method: EM (single particle) / Resolution: 3.4 Å

11570

7a5p

EMDB-11570: Human C Complex Spliceosome - MultiBody refined EXTENDED CORE
PDB-7a5p: Human C Complex Spliceosome - Medium-resolution PERIPHERY
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-11571:
Human C Complex Spliceosome - MultiBody refined BRR2/PRP16 region
Method: EM (single particle) / Resolution: 5.1 Å

EMDB-11572:
Human C Complex Spliceosome - MultiBody refined AQR/SYF1 region
Method: EM (single particle) / Resolution: 5.7 Å

EMDB-11573:
Human C Complex Spliceosome - MultiBody refined U2 Proteins region
Method: EM (single particle) / Resolution: 5.2 Å

EMDB-11574:
Human C Complex Spliceosome - MultiBody refined PRP19/Helical-Bundle region
Method: EM (single particle) / Resolution: 4.5 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-IHP:
INOSITOL HEXAKISPHOSPHATE / Phytic acid

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

ChemComp-ZN:
Unknown entry

Source
  • homo sapiens (human)
  • Human (human)
KeywordsSPLICING / human C complex / spliceosome / hC / human step 1 spliceosome

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