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TitleCryo-EM Structure of a Pre-catalytic Human Spliceosome Primed for Activation.
Journal, issue, pagesCell, Vol. 170, Issue 4, Page 701-713.e11, Year 2017
Publish dateAug 10, 2017
AuthorsKarl Bertram / Dmitry E Agafonov / Olexandr Dybkov / David Haselbach / Majety N Leelaram / Cindy L Will / Henning Urlaub / Berthold Kastner / Reinhard Lührmann / Holger Stark /
PubMed AbstractLittle is known about the spliceosome's structure before its extensive remodeling into a catalytically active complex. Here, we report a 3D cryo-EM structure of a pre-catalytic human spliceosomal B ...Little is known about the spliceosome's structure before its extensive remodeling into a catalytically active complex. Here, we report a 3D cryo-EM structure of a pre-catalytic human spliceosomal B complex. The U2 snRNP-containing head domain is connected to the B complex main body via three main bridges. U4/U6.U5 tri-snRNP proteins, which are located in the main body, undergo significant rearrangements during tri-snRNP integration into the B complex. These include formation of a partially closed Prp8 conformation that creates, together with Dim1, a 5' splice site (ss) binding pocket, displacement of Sad1, and rearrangement of Brr2 such that it contacts its U4/U6 substrate and is poised for the subsequent spliceosome activation step. The molecular organization of several B-specific proteins suggests that they are involved in negatively regulating Brr2, positioning the U6/5'ss helix, and stabilizing the B complex structure. Our results indicate significant differences between the early activation phase of human and yeast spliceosomes.
External linksCell / PubMed:28781166
MethodsEM (single particle)
Resolution4.5 - 9.9 Å
Structure data

3766

5o9z

EMDB-3766, PDB-5o9z:
Cryo-EM structure of a pre-catalytic human spliceosome primed for activation (B complex)
Method: EM (single particle) / Resolution: 4.5 Å

EMDB-3767:
Cryo-EM structure of a pre-catalytic human spliceosome primed for activation (B complex), 5.4A Map
Method: EM (single particle) / Resolution: 4.5 Å

EMDB-3768:
Cryo-EM structure of a pre-catalytic human spliceosome primed for activation (B complex). Unmasked refinement, sharpened map
Method: EM (single particle) / Resolution: 4.5 Å

EMDB-3769:
Cryo-EM structure of a pre-catalytic human spliceosome primed for activation (B complex). Unmasked refinement, NON-sharpened map
Method: EM (single particle) / Resolution: 9.9 Å

Source
  • homo sapiens (human)
  • Human (human)
KeywordsSPLICING / Spliceosome / pre-mRNA splicing / macromolecular complex

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