SUBSTITUTION OF PRE-MESSENGER RNA WITH PHOSPHOROTHIOATE LINKAGES REVEALS A NEW SPLICING-RELATED REACTION
- 5 September 1988
- journal article
- research article
- Vol. 263 (25) , 12295-12304
Abstract
In vitro transcription in the presence of a nucleoside 5''-O-(1-thiotriphosphate) has been used to prepare pre-mRNA analogues of the small intron of a rabbit .beta.-globin gene and flanking exon sequences. Incubation of transcripts prepared with adenosine 5''-O-(1-thiotriphosphate) in a HeLa cell nuclear extract showed that the presence of the thionucleotide in a transcript inhibited splicing, but a novel product was formed by cleavage three nucleotides upstream of the 3'' splice site. This product was formed with the same kinetics as the intermediates of a normal splicing reaction, and its formation depended on the presence of intact small nuclear RNAs U1, U2, and U6. We conclude that activation of 3'' splice site-proximal sequences need not be linked to exon ligation.This publication has 8 references indexed in Scilit:
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