Identification of sites of pre-MRNA/spliceosome association.
RNase H and synthetic DNA oligonucleotides were used to analyze the ribonucleoprotein (RNP) structure of the yeast spliceosome and to assay the pre-mRNA sequence requirements for step 1 of splicing. The data suggest that tight, stable contacts between the pre-mRNA and the spliceosome may be limited to the 5' splice site and branch point regions of the intron. A 30 nucleotide segment 3' of the branch point was found to be necessary for spliceosome maturation and essential for step 1 of splicing. Somewhat surprisingly, the 3' splice site was sensitive to nuclease digestion and completely dispensable for step 1 of splicing.
Morgan School of Biological Sciences, University of Kentucky, Lexington 40506-0225.
B C
BC
Rymond
D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure*; D004722:Endoribonucleases; D007438:Introns; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D016914:Ribonuclease H; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D009838:Oligodeoxyribonucleotides; D012322:RNA Precursors; D012333:RNA, Messenger; D004722:Endoribonucleases; D016914:Ribonuclease H