Human DDX17 unwinds rift valley fever virus non-coding RNAs

Corey R. Nelson, Tyler Mrozowich, Sean M. Park, Simmone D’souza, Amy Henrickson, Justin R.J. Vigar, Hans Joachim Wieden, Raymond J. Owens, Borries Demeler, Trushar R. Patel

Research output: Contribution to journalArticlepeer-review

18 Scopus citations


Rift 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. There-fore, 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 ultracen-trifugation 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 homoge-nous 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 regions.

Original languageEnglish
Article number54
Pages (from-to)1-17
Number of pages17
JournalInternational Journal of Molecular Sciences
Issue number1
StatePublished - Jan 1 2021


  • Analytical ultracen-trifuge
  • Fluorescent labeling
  • Helicase DDX17
  • Helicase assay
  • Host–viral interactions
  • Microscale thermophoresis
  • Rift Valley fever virus RNA
  • Small angle X-ray scattering


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