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Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry
Published on: January 17, 2025
In vitro binding of single-stranded RNA by human Dicer
Hemant K Kini1, S Patrick Walton
1Applied Biomolecular Engineering Laboratory of the Cellular and Biomolecular Laboratory, Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI 48824-1226, USA.
Abstract:
While Dicer alone has been shown to form stable complexes with double-stranded RNAs and short interfering RNAs, its interactions with single-stranded RNAs (ssRNAs) have not been characterized. Here, we show that recombinant human Dicer alone can bind 21-nt ssRNAs in vitro, independent of their sequence and structure. We also demonstrate that Dicer binds ssRNAs having a 5'-phosphate with greater affinity versus those with a 5'-hydroxyl. In addition, 3'-biotinylated ssRNAs are bound by Dicer with lower affinity than 3'-hydroxyl ssRNAs. The stability of ssRNA-Dicer complexes was found to depend on divalent cations. Together, our results suggest a role for the PAZ domain of Dicer in binding ssRNAs and may indicate roles for Dicer in cellular function beyond those currently known.
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