The RGG domain in the C-terminus of the DEAD box helicases Dbp2 and Ded1 is necessary for G-quadruplex destabilization
| Autor: | Kevin Kok-Phen Yan, Nasim Sabouri, Ikenna Obi |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
DEAD box
AcademicSubjects/SCI00010 Cell- och molekylärbiologi Cell Cycle Proteins 010402 general chemistry 01 natural sciences DEAD-box RNA Helicases 03 medical and health sciences chemistry.chemical_compound Protein Domains Schizosaccharomyces Genetics Humans Translation regulator activity 030304 developmental biology 0303 health sciences biology Nucleic Acid Enzymes C-terminus Biochemistry and Molecular Biology Membrane Proteins Helicase RNA biology.organism_classification RNA Helicase A 0104 chemical sciences Cell biology DNA-Binding Proteins G-Quadruplexes chemistry Schizosaccharomyces pombe biology.protein Schizosaccharomyces pombe Proteins RNA Helicases DNA Cell and Molecular Biology Biokemi och molekylärbiologi Protein Binding |
| Zdroj: | Nucleic Acids Research |
| Popis: | The identification of G-quadruplex (G4) binding proteins and insights into their mechanism of action are important for understanding the regulatory functions of G4 structures. Here, we performed an unbiased affinity-purification assay coupled with mass spectrometry and identified 30 putative G4 binding proteins from the fission yeast Schizosaccharomyces pombe. Gene ontology analysis of the molecular functions enriched in this pull-down assay included mRNA binding, RNA helicase activity, and translation regulator activity. We focused this study on three of the identified proteins that possessed putative arginine-glycine-glycine (RGG) domains, namely the Stm1 homolog Oga1 and the DEAD box RNA helicases Dbp2 and Ded1. We found that Oga1, Dbp2, and Ded1 bound to both DNA and RNA G4s in vitro. Both Dbp2 and Ded1 bound to G4 structures through the RGG domain located in the C-terminal region of the helicases, and point mutations in this domain weakened the G4 binding properties of the helicases. Dbp2 and Ded1 destabilized less thermostable G4 RNA and DNA structures, and this ability was independent of ATP but dependent on the RGG domain. Our study provides the first evidence that the RGG motifs in DEAD box helicases are necessary for both G4 binding and G4 destabilization. |
| Databáze: | OpenAIRE |
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