A Specific RNA Hairpin Loop Structure Binds the RNA Recognition Motifs of the Drosophila SR Protein B52
| Autor: | John T. Lis, Bryan E. Hoffman, Hua Shi |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
RNA Splicing
Molecular Sequence Data RNA-binding protein Biology Polymerase Chain Reaction Structure-Activity Relationship SR protein RNA Precursors Animals Drosophila Proteins Molecular Biology Genetics Base Sequence Intron Nuclear Proteins RNA-Binding Proteins RNA Cell Biology Phosphoproteins Non-coding RNA Cell biology Drosophila melanogaster RNA editing RNA splicing Nucleic Acid Conformation RNA Splicing Factors Small nuclear RNA Research Article |
| Zdroj: | Molecular and Cellular Biology. 17:2649-2657 |
| ISSN: | 1098-5549 |
| DOI: | 10.1128/mcb.17.5.2649 |
| Popis: | B52, also known as SRp55, is a member of the Drosophila melanogaster SR protein family, a group of nuclear proteins that are both essential splicing factors and specific splicing regulators. Like most SR proteins, B52 contains two RNA recognition motifs in the N terminus and a C-terminal domain rich in serine-arginine dipeptide repeats. Since B52 is an essential protein and is expected to play a role in splicing a subset of Drosophila pre-mRNAs, its function is likely to be mediated by specific interactions with RNA. To investigate the RNA-binding specificity of B52, we isolated B52-binding RNAs by selection and amplification from a pool of random RNA sequences by using full-length B52 protein as the target. These RNAs contained a conserved consensus motif that constitutes the core of a secondary structural element predicted by energy minimization. Deletion and substitution mutations defined the B52-binding site on these RNAs as a hairpin loop structure covering about 20 nucleotides, which was confirmed by structure-specific enzymatic probing. Finally, we demonstrated that both RNA recognition motifs of B52 are required for RNA binding, while the RS domain is not involved in this interaction. |
| Databáze: | OpenAIRE |
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