Structural basis for recognition of intron branchpoint RNA by yeast Msl5 and selective effects of interfacial mutations on splicing of yeast pre-mRNAs Journal Article


Authors: Jacewicz, A.; Chico, L.; Smith, P.; Schwer, B.; Shuman, S.
Article Title: Structural basis for recognition of intron branchpoint RNA by yeast Msl5 and selective effects of interfacial mutations on splicing of yeast pre-mRNAs
Abstract: Saccharomyces cerevisiae Msl5 orchestrates spliceosome assembly by binding the intron branchpoint sequence 5' -UACUAAC and, with its heterodimer partner protein Mud2, establishing cross intron-bridging interactions with the U1 snRNP at the 5' splice site. Here we define the central Msl5 KH-QUA2 domain as sufficient for branchpoint RNA recognition. The 1.8 Å crystal structure of Msl5-(KH-QUA2) bound to the branchpoint highlights an extensive network of direct and water-mediated protein-RNA and intra-RNA atomic contacts at the interface that illuminate how Msl5 recognizes each nucleobase of the UACUAAC element. The Msl5 structure rationalizes a large body of mutational data and inspires new functional studies herein, which reveal how perturbations of the Msl5•RNA interface impede the splicing of specific yeast pre-mRNAs. We also identify interfacial mutations in Msl5 that bypass the essentiality of Sub2, a DExD-box ATPase implicated in displacing Msl5 from the branchpoint in exchange for the U2 snRNP. These studies establish an atomic resolution framework for understanding splice site selection and early spliceosome dynamics.
Keywords: saccharomyces cerevisiae; branchpoint binding protein; spliceosome assembly; kh-qua2 domain
Journal Title: RNA
Volume: 21
Issue: 3
ISSN: 1355-8382
Publisher: Cold Spring Harbor Laboratory Press  
Date Published: 2015-03-01
Start Page: 401
End Page: 414
Language: English
DOI: 10.1261/rna.048942.114
PROVIDER: scopus
PMCID: PMC4338336
PUBMED: 25587180
DOI/URL:
Notes: Export Date: 2 March 2015 -- Source: Scopus
Altmetric
Citation Impact
MSK Authors
  1. Stewart H Shuman
    514 Shuman