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In vitro binding of single‐stranded RNA by human Dicer
Author(s) -
Kini Hemant K.,
Walton S. Patrick
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.11.010
Subject(s) - dicer , chemistry , rna , ribonuclease iii , small interfering rna , in vitro , recombinant dna , microbiology and biotechnology , biophysics , biochemistry , biology , rna interference , gene
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.