Reference: Tardiff DF and Rosbash M (2006) Arrested yeast splicing complexes indicate stepwise snRNP recruitment during in vivo spliceosome assembly. RNA 12(6):968-79

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Abstract


Pre-mRNA splicing is catalyzed by the spliceosome, a macromolecular machine dedicated to intron removal and exon ligation. Despite an abundance of in vitro information and a small number of in vivo studies, the pathway of yeast (Saccharomyces cerevisiae) in vivo spliceosome assembly remains uncertain. To address this situation, we combined in vivo depletions of U1, U2, or U5 snRNAs with chromatin immunoprecipitation (ChIP) analysis of other splicing snRNPs along an intron-containing gene. The data indicate that snRNP recruitment to nascent pre-mRNA predominantly proceeds via the canonical three-step assembly pathway: first U1, then U2, and finally the U4/U6*U5 tri-snRNP. Tandem affinity purification (TAP) using a U2 snRNP-tagged protein allowed the characterization of in vivo assembled higher-order splicing complexes. Consistent with an independent snRNP assembly pathway, we observed high levels of U1-U2 prespliceosomes under U5-depletion conditions, and we observed significant levels of a U2/U5/U6/Prp19-complex mature splicing complex under wild-type conditions. These complexes have implications for the steady-state distribution of snRNPs within nuclei and also reinforce the stepwise recruitment of U1, U2, and the tri-snRNP during in vivo spliceosome assembly.

Reference Type
Journal Article | Research Support, N.I.H., Extramural
Authors
Tardiff DF, Rosbash M
Primary Lit For
PRP21 | LUC7 | SMX2 | SMD1 | MUD1 | SNP1 | PRP40 | PRP8 | PRP42 | SNU56 | MSL1 | ... Show all
Additional Lit For
ISY1 | PRP45 | SNU114 | CWC2 | SYF1 | SNR6 | IST3 | CWC15 | BRR2 | CEF1 | ECM2 | ... Show all

Gene Ontology Annotations


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Gene/Complex Qualifier Gene Ontology Term Aspect Annotation Extension Evidence Method Source Assigned On Reference

Interaction Annotations


Genetic Interactions

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Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

Physical Interactions 40 entries for 40 genes

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InteractorInteractorAssayAnnotationActionModification
BRR2LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
CEF1LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
CLF1LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
CUS1LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
CWC15LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
CWC2LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
ECM2LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
HSH155LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
HSH49LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
ISY1LEA1Affinity Capture-MSmanually curatedHit-BaitNo Modification
Showing 1 to 10 of 40 entries