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TitleCryo-EM analyses of dimerized spliceosomes provide new insights into the functions of B complex proteins.
Journal, issue, pagesEMBO J, Vol. 43, Issue 6, Page 1065-1088, Year 2024
Publish dateFeb 21, 2024
AuthorsZhenwei Zhang / Vinay Kumar / Olexandr Dybkov / Cindy L Will / Henning Urlaub / Holger Stark / Reinhard Lührmann /
PubMed AbstractThe B complex is a key intermediate stage of spliceosome assembly. To improve the structural resolution of monomeric, human spliceosomal B (hB) complexes and thereby generate a more comprehensive hB ...The B complex is a key intermediate stage of spliceosome assembly. To improve the structural resolution of monomeric, human spliceosomal B (hB) complexes and thereby generate a more comprehensive hB molecular model, we determined the cryo-EM structure of B complex dimers formed in the presence of ATP S. The enhanced resolution of these complexes allows a finer molecular dissection of how the 5' splice site (5'ss) is recognized in hB, and new insights into molecular interactions of FBP21, SNU23 and PRP38 with the U6/5'ss helix and with each other. It also reveals that SMU1 and RED are present as a heterotetrameric complex and are located at the interface of the B dimer protomers. We further show that MFAP1 and UBL5 form a 5' exon binding channel in hB, and elucidate the molecular contacts stabilizing the 5' exon at this stage. Our studies thus yield more accurate models of protein and RNA components of hB complexes. They further allow the localization of additional proteins and protein domains (such as SF3B6, BUD31 and TCERG1) whose position was not previously known, thereby uncovering new functions for B-specific and other hB proteins during pre-mRNA splicing.
External linksEMBO J / PubMed:38383864 / PubMed Central
MethodsEM (single particle)
Resolution3.1 - 15.0 Å
Structure data

EMDB-18225, PDB-8q7n:
cryo-EM structure of the human spliceosomal B complex protomer (tri-snRNP core region)
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-18529, PDB-8qo9:
Cryo-EM structure of a human spliceosomal B complex protomer
Method: EM (single particle) / Resolution: 5.29 Å

EMDB-19062: BRR2 part of the human spliceosomal B complex
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-19063: cryo-EM structure of a dimerized human spliceosomal B complex
Method: EM (single particle) / Resolution: 15.0 Å

Source
  • homo sapiens (human)
KeywordsSPLICING / spliceosome / pre-catalytic spliceosome / spliceosomal B complex

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