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Structure paper

TitleMechanism of 5' splice site transfer for human spliceosome activation.
Journal, issue, pagesScience, Vol. 364, Issue 6438, Page 362-367, Year 2019
Publish dateApr 26, 2019
AuthorsClément Charenton / Max E Wilkinson / Kiyoshi Nagai /
PubMed AbstractThe prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5' splice site (5'SS) and branch point sequence, associates with the U4/U6.U5 ...The prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5' splice site (5'SS) and branch point sequence, associates with the U4/U6.U5 tri-snRNP to form the fully assembled precatalytic pre-B spliceosome. Here, we report cryo-electron microscopy structures of the human pre-B complex captured before U1 snRNP dissociation at 3.3-angstrom core resolution and the human tri-snRNP at 2.9-angstrom resolution. U1 snRNP inserts the 5'SS-U1 snRNA helix between the two RecA domains of the Prp28 DEAD-box helicase. Adenosine 5'-triphosphate-dependent closure of the Prp28 RecA domains releases the 5'SS to pair with the nearby U6 ACAGAGA-box sequence presented as a mobile loop. The structures suggest that formation of the 5'SS-ACAGAGA helix triggers remodeling of an intricate protein-RNA network to induce Brr2 helicase relocation to its loading sequence in U4 snRNA, enabling Brr2 to unwind the U4/U6 snRNA duplex to allow U6 snRNA to form the catalytic center of the spliceosome.
External linksScience / PubMed:30975767 / PubMed Central
MethodsEM (single particle)
Resolution2.9 - 28.0 Å
Structure data

4658

6qw6

EMDB-4658, PDB-6qw6:
Structure of the human U5.U4/U6 tri-snRNP at 2.9A resolution.
Method: EM (single particle) / Resolution: 2.92 Å

4665

6qx9

EMDB-4665, PDB-6qx9:
Structure of a human fully-assembled precatalytic spliceosome (pre-B complex).
Method: EM (single particle) / Resolution: 3.28 Å

EMDB-4672:
Human U5.U4/U6 tri-snRNP, multi-body refinement, Brr2 map
Method: EM (single particle) / Resolution: 3.11 Å

EMDB-4673:
Human U5.U4/U6 tri-snRNP, multi-body refinement, core map
Method: EM (single particle) / Resolution: 2.9 Å

EMDB-4674:
Human U5.U4/U6 tri-snRNP, multi-body refinement, foot map
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-4675:
Human U5.U4/U6 tri-snRNP, multi-body refinement, U4 core map
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-4676:
Human U5.U4/U6 tri-snRNP, multi-body refinement, Prp4 map
Method: EM (single particle) / Resolution: 3.38 Å

EMDB-4686:
Human U5.U4/U6 tri-snRNP, refined after focused classification on Prp28
Method: EM (single particle) / Resolution: 2.98 Å

EMDB-4687:
Human fully assembled spliceosome (pre-B complex), overall map with stable U1 snRNP
Method: EM (single particle) / Resolution: 6.45 Å

EMDB-4688:
Human fully assembled spliceosome (pre-B complex), core map after focused classification on Prp4 kinase
Method: EM (single particle) / Resolution: 6.19 Å

EMDB-4689:
Human fully assembled spliceosome (pre-B complex), focused refinement on U1 snRNP
Method: EM (single particle) / Resolution: 28.0 Å

EMDB-4690:
Human fully assembled spliceosome (pre-B complex), focused refinement on SF3B
Method: EM (single particle) / Resolution: 27.0 Å

Chemicals

ChemComp-IHP:
INOSITOL HEXAKISPHOSPHATE

ChemComp-MG:
Unknown entry

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

ChemComp-ZN:
Unknown entry

Source
  • homo sapiens (human)
  • Human (human)
  • human adenovirus 2
KeywordsSPLICING / RNP complex / RNA / protein / spliceosome

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