Substitution of pre-mRNA with phosphorothioate linkages reveals a new splicing-related reaction
| Autor: | A D, Griffiths, B V, Potter, I C, Eperon |
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| Rok vydání: | 1988 |
| Předmět: |
Exonucleases
Base Sequence Transcription Genetic RNA Splicing Molecular Sequence Data Affinity Labels DNA-Directed RNA Polymerases Exons Thionucleotides Introns Globins Adenosine Triphosphate RNA Small Nuclear Endoribonucleases RNA Precursors Animals Humans Electrophoresis Polyacrylamide Gel T-Phages RNA Messenger Rabbits HeLa Cells |
| Zdroj: | The Journal of biological chemistry. 263(25) |
| ISSN: | 0021-9258 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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