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TitleMechanism of protein-guided folding of the active site U2/U6 RNA during spliceosome activation.
Journal, issue, pagesScience, Vol. 370, Issue 6523, Year 2020
Publish dateDec 18, 2020
AuthorsCole Townsend / Majety N Leelaram / Dmitry E Agafonov / Olexandr Dybkov / Cindy L Will / Karl Bertram / Henning Urlaub / Berthold Kastner / Holger Stark / Reinhard Lührmann /
PubMed AbstractSpliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway ...Spliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway of the latter and the mechanism whereby proteins aid its proper folding. Here, we report the cryo-electron microscopy structures of two human, activated spliceosome precursors (that is, pre-B complexes) at core resolutions of 3.9 and 4.2 angstroms. These structures elucidate the order of the numerous protein exchanges that occur during activation, the mutually exclusive interactions that ensure the correct order of ribonucleoprotein rearrangements needed to form the U2/U6 catalytic RNA, and the stepwise folding pathway of the latter. Structural comparisons with mature B complexes reveal the molecular mechanism whereby a conformational change in the scaffold protein PRP8 facilitates final three-dimensional folding of the U2/U6 catalytic RNA.
External linksScience / PubMed:33243851
MethodsEM (single particle)
Resolution3.9 - 8.0 Å
Structure data

11693

7aav

EMDB-11693, PDB-7aav:
Human pre-Bact-2 spliceosome core structure
Method: EM (single particle) / Resolution: 4.2 Å

11694

7abf

EMDB-11694, PDB-7abf:
Human pre-Bact-1 spliceosome core structure
Method: EM (single particle) / Resolution: 3.9 Å

11695

7abg

EMDB-11695, PDB-7abg:
Human pre-Bact-1 spliceosome
Method: EM (single particle) / Resolution: 7.8 Å

11696

7abh

EMDB-11696, PDB-7abh:
Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
Method: EM (single particle) / Resolution: 4.5 Å

11697

7abi

EMDB-11697, PDB-7abi:
Human pre-Bact-2 spliceosome
Method: EM (single particle) / Resolution: 8.0 Å

Chemicals

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

ChemComp-MG:
Unknown entry

ChemComp-KGN:
D-chiro inositol hexakisphosphate

ChemComp-IHP:
INOSITOL HEXAKISPHOSPHATE / Phytic acid

ChemComp-GTG:
7-METHYL-GUANOSINE-5'-TRIPHOSPHATE-5'-GUANOSINE

Source
  • homo sapiens (human)
  • synthetic construct (others)
  • Human (human)
KeywordsSPLICING / Complex / spliceosome / catalytic activation

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