Yeast mRNA splicing in vitro.
Synthetic actin and CYH2 pre-mRNAs containing a single intron are accurately spliced in a soluble whole cell extract of yeast. Splicing in vitro requires ATP. The excised intron is released as a lariat in which an RNA branch connects the 5' end of the molecule to the last A in the "intron conserved sequence" UACUAAC. Two other discrete RNA species produced during splicing in vitro may represent reaction intermediates: free, linear exon 1 and a form of the intron lariat extending beyond the 3' splice site to include exon 2. Both lariat forms correspond to molecules previously shown to be produced during yeast pre-mRNA splicing in vivo.
D000199:Actins / Q000235:genetics*; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D005796:Genes; D005800:Genes, Fungal; D009693:Nucleic Acid Hybridization; D009694:Nucleic Acid Precursors / Q000235:genetics; D010957:Plasmids; D012322:RNA Precursors; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics*; D006163:Ribonuclease T1; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014158:Transcription, Genetic
D000199:Actins; D009694:Nucleic Acid Precursors; D012322:RNA Precursors; D012333:RNA, Messenger; D004262:DNA Restriction Enzymes; D006163:Ribonuclease T1