Sequence-specific affinity selection of mammalian splicing complexes.
Antisense oligonucleotides made of 2'-OMe RNA are shown to bind specifically and efficiently to targeted sites on pre-mRNA substrates, allowing affinity selection of splicing complexes using streptavidin/biotin chromatography. The position of probe binding to the pre-mRNA influences which type of splicing complex can be selected. The accessibility of pre-mRNA sequences to antisense probes changes during the course of the splicing reaction. U1, U2, U4, U5 and U6 snRNAs are all detected in affinity-selected mammalian splicing complexes. However, antisense oligonucleotides targeted to snRNAs can block the binding of specific snRNPs to pre-mRNA. Quantitative affinity selection analyses show that only a small fraction of snRNPs in a HeLa nuclear splicing extract participate in spliceosome formation.
Ryder
U
U
European Molecular Biology Laboratory, Heidelberg, FRG.
Sproat
B S
BS
Lamond
A I
AI
D001483:Base Sequence; D002846:Chromatography, Affinity; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009843:Oligoribonucleotides / Q000378:metabolism; D012322:RNA Precursors / Q000378:metabolism; D012326:RNA Splicing; D016372:RNA, Antisense / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear
D009843:Oligoribonucleotides; D012322:RNA Precursors; D016372:RNA, Antisense; D012342:RNA, Small Nuclear; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear