U1 snRNP-ASF/SF2 interaction and 5' splice site recognition: characterization of required elements
| Autor: | Cindy L. Will, Jin Wang, James L. Manley, Zvi Pasman, Reinhard Lührmann, Mariano A. Garcia-Blanco, Sharon F. Jamison |
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| Jazyk: | angličtina |
| Rok vydání: | 1995 |
| Předmět: |
Genetics
Spliceosome Binding Sites Base Sequence Serine-Arginine Splicing Factors RNA Splicing Molecular Sequence Data Nuclear Proteins RNA-Binding Proteins RNA-binding protein Biology Non-coding RNA Cell biology Ribonucleoprotein U1 Small Nuclear SR protein RNA splicing RNA Precursors Spliceosomes Animals Humans splice snRNP Binding domain Protein Binding |
| Popis: | Members of the SR family of proteins, can collaborate with U1 snRNP in the recognition of 5' splice sites in pre-messenger RNAs. We have previously shown that purified U1 snRNP and ASF/SF2 form a ternary complex with pre-mRNA, which is dependent on a functional 5' splice site. In this manuscript we dissect the requirements for the formation of this complex. Sequences in the pre-mRNA, domains in ASF/SF2 and components of the U1 snRNP particle are shown to be required for complex formation. We had shown that sequences at the 5' splice site of PIP7. A are necessary and now we show these are sufficient for complex formation. Furthermore, we show that one functional RNA binding domain and the RS domain are both required for ASF/SF2 to participate in complex formation. The RNA binding domains were redundant in this assay, suggesting that either domain can interact with the pre-messenger RNA. Finally, our experiments show no function for the U1-specific A protein in complex formation, whereas a function for U1-specific C protein was strongly suggested. The study of the earliest interactions between pre-mRNA and splicing factors suggests a model for 5' splice site recognition. |
| Databáze: | OpenAIRE |
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