Pre-mRNA splicing is executed by the spliceosome, which has eight major functional states each with distinct composition. Five of these eight human spliceosomal complexes, all preceding exon ligation, have been structurally characterized. In this study, we report the cryo-electron microscopy structures of the human post-catalytic spliceosome (P complex) and intron lariat spliceosome (ILS) at average resolutions of 3.0 and 2.9 Å, respectively. In the P complex, the ligated exon remains anchored to loop I of U5 small nuclear RNA, and the 3'-splice site is recognized by the junction between the 5'-splice site and the branch point sequence. The ATPase/helicase Prp22, along with the ligated exon and eight other proteins, are dissociated in the P-to-ILS transition. Intriguingly, the ILS complex exists in two distinct conformations, one with the ATPase/helicase Prp43 and one without. Comparison of these three late-stage human spliceosomes reveals mechanistic insights into exon release and spliceosome disassembly.
Mutations in a yeast intron demonstrate the importance of specific conserved nucleotides for the two stages of nuclear mRNA splicing
Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei
Interaction between the first and last nucleotides of pre-mRNA introns is a determinant of 3' splice site selection in S. cerevisiae
Evidence for an essential non-Watson-Crick interaction between the first and last nucleotides of a nuclear pre-mRNA intron
The role of branchpoint-3' splice site spacing and interaction between intron terminal nucleotides in 3' splice site selection in Saccharomyces cerevisiae
Divergent effects of chaperone overexpression and ethanol supplementation on inclusion body formation in recombinant Escherichia coli
Prp43 is an essential RNA-dependent ATPase required for release of lariat-intron from the spliceosome.
Purification and characterization of native spliceosomes suitable for three-dimensional structural analysis
Optimal determination of particle orientation, absolute hand, and contrast loss in single-particle electron cryomicroscopy
Automated acquisition of cryo-electron micrographs for single particle reconstruction on an FEI Tecnai electron microscope
Opposing classes of prp8 alleles modulate the transition between the catalytic steps of pre-mRNA splicing
Isolation of XAB2 complex involved in pre-mRNA splicing, transcription, and transcription-coupled repair
Cwc2 and its human homologue RBM22 promote an active conformation of the spliceosome catalytic centre
Human CWC22 escorts the helicase eIF4AIII to spliceosomes and promotes exon junction complex assembly
Human spliceosomal protein CWC22 plays a role in coupling splicing to exon junction complex deposition and nonsense-mediated decay
Structural basis of Brr2-Prp8 interactions and implications for U5 snRNP biogenesis and the spliceosome active site
High-resolution noise substitution to measure overfitting and validate resolution in 3D structure determination by single particle electron cryomicroscopy
Atomic-accuracy models from 4.5-Å cryo-electron microscopy data with density-guided iterative local refinement
The crystal structure of human DEAH-box RNA helicase 15 reveals a domain organization of the mammalian DEAH/RHA family
Cryo-electron microscopy snapshots of the spliceosome: structural insights into a dynamic ribonucleoprotein machine
All-atom simulations disentangle the functional dynamics underlying gene maturation in the intron lariat spliceosome
Termination of pre-mRNA splicing requires that the ATPase and RNA unwindase Prp43p acts on the catalytic snRNA U6
The inactive C-terminal cassette of the dual-cassette RNA helicase BRR2 both stimulates and inhibits the activity of the N-terminal helicase unit.
Structural and functional insights into CWC27/CWC22 heterodimer linking the exon junction complex to spliceosomes
Mutations in Spliceosomal Genes PPIL1 and PRP17 Cause Neurodegenerative Pontocerebellar Hypoplasia with Microcephaly.
Alternative splicing a regulated gene expression process that allows a single genetic sequence to code for multiple proteins. Here is that latest research.