The glucocorticoid receptor DNA-binding domain recognizes RNA hairpin structures with high affinity
| Autor: | Nickolaus C Lammer, Zachariah E Holmes, Deborah S. Wuttke, Nicholas V. Parsonnet, Robert T. Batey |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular Plasma protein binding Biology Binding Competitive 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Receptors Glucocorticoid Protein Domains Genetics RNA and RNA-protein complexes Humans Binding site 030304 developmental biology 0303 health sciences Binding Sites Base Sequence RNA DNA-binding domain DNA Non-coding RNA DNA-Binding Proteins RNA silencing chemistry Biophysics Nucleic Acid Conformation GAS5 030217 neurology & neurosurgery Protein Binding |
| Popis: | The glucocorticoid receptor (GR) binds the noncoding RNA Gas5 via its DNA-binding domain (DBD) with functional implications in pro-apoptosis signaling. Here, we report a comprehensive in vitro binding study where we have determined that GR-DBD is a robust structure-specific RNA-binding domain. GR-DBD binds to a diverse range of RNA hairpin motifs, both synthetic and biologically derived, with apparent mid-nanomolar affinity while discriminating against uniform dsRNA. As opposed to dimeric recognition of dsDNA, GR-DBD binds to RNA as a monomer and confers high affinity primarily through electrostatic contacts. GR-DBD adopts a discrete RNA-bound state, as assessed by NMR, distinct from both free and DNA-bound. NMR and alanine mutagenesis suggest a heightened involvement of the C-terminal α-helix of the GR-DBD in RNA-binding. RNA competes for binding with dsDNA and occurs in a similar affinity range as dimer binding to the canonical DNA element. Given the prevalence of RNA hairpins within the transcriptome, our findings strongly suggest that many RNAs have potential to impact GR biology. |
| Databáze: | OpenAIRE |
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