Nucleic acids research, 2017-01, Vol.45 (1), p.206-214
2017
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- Autor(en) / Beteiligte
- Titel
- RHAU helicase stabilizes G4 in its nucleotide-free state and destabilizes G4 upon ATP hydrolysis
- Ist Teil von
- Nucleic acids research, 2017-01, Vol.45 (1), p.206-214
- Ort / Verlag
- England
- Erscheinungsjahr
- 2017
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- The DEAH-box ATP-dependent RHAU helicases specifically unfold RNA and DNA G-quadruplexes (G4s). However, it remains unclear how the RHAU's G4 unfolding activity is coupled to different stages of the ATPase cycle. Here, using a single-molecule manipulation approach, we show that binding of Drosophila RHAU stabilizes an intramolecularly folded parallel DNA G4 against mechanical unfolding in its nucleotide-free and in its AMP-PNP or ADP bound states, while it destabilizes the G4 when coupled to ATP hydrolysis. Importantly, our results show that the ADP·AlF[Formula: see text]-bound RHAU does not stabilize the G4. We also found that both a single-stranded 3' DNA tail and the RSM domain of RHAU that binds specifically to the G4 structure, are dispensable for the stabilization of the G4, but both are required for G4 destabilization. Our study provides the first evidence that the unfolding kinetics of a G-quadruplex can be modulated by different nucleotide-bound states of the helicase.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0305-1048eISSN: 1362-4962DOI: 10.1093/nar/gkw881Titel-ID: cdi_pubmed_primary_28069994
- Format
- –
- Schlagworte
- Adenosine Diphosphate - chemistry, Adenosine Diphosphate - genetics, Adenosine Diphosphate - metabolism, Adenosine Monophosphate - chemistry, Adenosine Monophosphate - genetics, Adenosine Monophosphate - metabolism, Adenosine Triphosphate - chemistry, Adenosine Triphosphate - genetics, Adenosine Triphosphate - metabolism, Animals, Cloning, Molecular, DEAD-box RNA Helicases - chemistry, DEAD-box RNA Helicases - genetics, DEAD-box RNA Helicases - metabolism, DNA - chemistry, DNA - genetics, DNA - metabolism, Drosophila melanogaster - chemistry, Drosophila melanogaster - metabolism, Drosophila Proteins - chemistry, Drosophila Proteins - genetics, Drosophila Proteins - metabolism, Escherichia coli - genetics, Escherichia coli - metabolism, G-Quadruplexes, Gene Expression, Humans, Hydrolysis, Isoenzymes - chemistry, Isoenzymes - genetics, Isoenzymes - metabolism, Kinetics, Magnetic Fields, Optical Tweezers, Protein Binding, Protein Domains, Recombinant Proteins - chemistry, Recombinant Proteins - genetics, Recombinant Proteins - metabolism, RNA - chemistry, RNA - genetics, RNA - metabolism, Single Molecule Imaging, Thermodynamics