Sequence requirements in different steps of the pre-mRNA splicing reaction: analysis by the RNA modification-exclusion technique.
The stepwise assembly of splicing complexes and the subsequent splicing reaction were analyzed by the RNA modification-exclusion technique, which generates the equivalent of a complete set of point mutations in a single reaction. We found that although the sequences surrounding the 5' splice site, the branch point, and the 3' splice site, including the 3' AG, were required for presplicing complex formation, modified nucleotides at these positions were not completely excluded. The same sequences were required for splicing complex formation; however, modified nucleotides in these sequences were excluded to a much greater extent.
Department of Cell Biology, Biocenter of the University of Basel, Switzerland.
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Lang
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Keller
D000260:Adenoviruses, Human / Q000235:genetics; D001483:Base Sequence; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D012322:RNA Precursors / Q000235:genetics* / Q000302:isolation & purification; D012326:RNA Splicing; D014158:Transcription, Genetic
D015345:Oligonucleotide Probes; D012322:RNA Precursors