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Molecular Biology Reagents> | We synthesized this mutant region through Genscript and replaced the 5′ SS-to-BPS region of the ACT1–CUP1 reporter plasmid19 with this sequence to generate the mutant reporter plasmid pMA6. | Get A Quote |
The molecular mechanisms of exon definition and back-splicing are fundamental unanswered questions in pre-mRNA splicing. Here we report cryo-electron microscopy structures of the yeast spliceosomal E complex assembled on introns, providing a view of the earliest event in the splicing cycle that commits pre-mRNAs to splicing. The E complex architecture suggests that the same spliceosome can assemble across an exon, and that it either remodels to span an intron for canonical linear splicing (typically on short exons) or catalyses back-splicing to generate circular RNA (on long exons). The model is supported by our experiments, which show that an E complex assembled on the middle exon of yeast EFM5 or HMRA1 ... More