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• W2100065623 endingPage "412" @default.
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• W2100065623 abstract "In spliceosomes, dynamic RNA/RNA and RNA/protein interactions position the pre-mRNA substrate for the two chemical steps of splicing. Not all of these interactions have been characterized, in part because it has not been possible to arrest the complex at clearly defined states relative to chemistry. Previously, it was shown in yeast that the DEAD/H-box protein Prp22 requires an extended 3' exon to promote mRNA release from the spliceosome following second-step chemistry. In line with that observation, we find that shortening the 3' exon blocks cleaved lariat intron and mRNA release in human splicing extracts, which allowed us to stall human spliceosomes in a new post-catalytic complex (P complex). In comparison to C complex, which is blocked at a point following first-step chemistry, we detect specific differences in RNA substrate interactions near the splice sites. These differences include extended protection across the exon junction and changes in protein crosslinks to specific sites in the 5' and 3' exons. Using selective reaction monitoring (SRM) mass spectrometry, we quantitatively compared P and C complex proteins and observed enrichment of SF3b components and loss of the putative RNA-dependent ATPase DHX35. Electron microscopy revealed similar structural features for both complexes. Notably, additional density is present when complexes are chemically fixed, which reconciles our results with previously reported C complex structures. Our ability to compare human spliceosomes before and after second-step chemistry has opened a new window to rearrangements near the active site of spliceosomes, which may play roles in exon ligation and mRNA release." @default.
• W2100065623 created "2016-06-24" @default.
• W2100065623 creator A5029492866 @default.
• W2100065623 creator A5035358710 @default.
• W2100065623 creator A5047624012 @default.
• W2100065623 creator A5068865174 @default.
• W2100065623 date "2013-01-23" @default.
• W2100065623 modified "2023-09-25" @default.
• W2100065623 title "Rearrangements within human spliceosomes captured after exon ligation" @default.
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• W2100065623 doi "https://doi.org/10.1261/rna.034223.112" @default.
• W2100065623 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3677250" @default.
• W2100065623 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/23345524" @default.
• W2100065623 hasPublicationYear "2013" @default.
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