Human DDX17 unwinds Rift Valley fever virus non-coding RNAs

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dc.contributor.authorNelson, Corey R.
dc.contributor.authorMrozowich, Tyler
dc.contributor.authorPark, Sean M.
dc.contributor.authorD'Souza, Simmone
dc.contributor.authorHenrickson, Amy
dc.contributor.authorVigar, Justin R. J.
dc.contributor.authorWieden, Hans-Joachim
dc.contributor.authorOwens, Raymond J.
dc.contributor.authorDemeler, Borries
dc.contributor.authorPatel, Trushar R.
dc.date.accessioned2021-07-30T21:01:02Z
dc.date.available2021-07-30T21:01:02Z
dc.date.issued2020
dc.descriptionOpen access article. Creative Commons Attribution 4.0 International License (CC BY 4.0) appliesen_US
dc.description.abstractRift Valley fever virus (RVFV) is a mosquito-transmitted virus from the Bunyaviridae family that causes high rates of mortality and morbidity in humans and ruminant animals. Previous studies indicated that DEAD-box helicase 17 (DDX17) restricts RVFV replication by recognizing two primary non-coding RNAs in the S-segment of the genome: the intergenic region (IGR) and 5′ non-coding region (NCR). However, we lack molecular insights into the direct binding of DDX17 with RVFV non-coding RNAs and information on the unwinding of both non-coding RNAs by DDX17. Therefore, we performed an extensive biophysical analysis of the DDX17 helicase domain (DDX17135–555) and RVFV non-coding RNAs, IGR and 5’ NCR. The homogeneity studies using analytical ultracentrifugation indicated that DDX17135–555, IGR, and 5’ NCR are pure. Next, we performed small-angle X-ray scattering (SAXS) experiments, which suggested that DDX17 and both RNAs are homogenous as well. SAXS analysis also demonstrated that DDX17 is globular to an extent, whereas the RNAs adopt an extended conformation in solution. Subsequently, microscale thermophoresis (MST) experiments were performed to investigate the direct binding of DDX17 to the non-coding RNAs. The MST experiments demonstrated that DDX17 binds with the IGR and 5’ NCR with a dissociation constant of 5.77 ± 0.15 µM and 9.85 ± 0.11 µM, respectively. As DDX17135–555 is an RNA helicase, we next determined if it could unwind IGR and NCR. We developed a helicase assay using MST and fluorescently-labeled oligos, which suggested DDX17135–555 can unwind both RNAs. Overall, our study provides direct evidence of DDX17135–555 interacting with and unwinding RVFV non-coding regionsen_US
dc.description.peer-reviewYesen_US
dc.description.sponsorshipOpen access article. Creative Commons Attribution 4.0 International License (CC BY 4.0) appliesen_US
dc.identifier.citationNelson, C. R., Mrozowich, T., Park, S. M., D'souza, S., Henrickson, A., Vigar, J.R. J., Wieden, H.-J., Owens, R. J., Demeler, B., & Patel, T. R. (2020). Human DDX17 unwinds Rift Valley fever virus non-coding RNAs. International Journal of Molecular Sciences, 22(1), Article 54. https://doi.org/10.3390/ijms22010054en_US
dc.identifier.urihttps://hdl.handle.net/10133/5981
dc.language.isoen_USen_US
dc.publisherMDPIen_US
dc.publisher.departmentDepartment of Chemistry and Biochemistryen_US
dc.publisher.facultyArts and Scienceen_US
dc.publisher.institutionUniversity of Lethbridgeen_US
dc.publisher.institutionUniversity of Calgaryen_US
dc.publisher.institutionRutherford Appleton Laboratoriesen_US
dc.publisher.institutionUniversity of Montanaen_US
dc.publisher.institutionUniversity of Albertaen_US
dc.publisher.urlhttps://doi.org/10.3390/ijms22010054en_US
dc.subjectRift Valley fever virus RNAen_US
dc.subjectHelicase DDX17en_US
dc.subjectHost-viral interactionsen_US
dc.subjectAnalytical ultracentrifugeen_US
dc.subjectMicroscale thermophoresisen_US
dc.subjectFluorescent labelingen_US
dc.subjectHelicase assayen_US
dc.subject.lcshDNA helicases
dc.subject.lcshHost-virus relationships
dc.subject.lcshCentrifuges
dc.subject.lcshSmall-angle x-ray scattering
dc.titleHuman DDX17 unwinds Rift Valley fever virus non-coding RNAsen_US
dc.typeArticleen_US
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Wieden, Hans-Joachim
Demeler, Borries
Patel, Trushar