In vitro binding of single-stranded RNA by human Dicer

作者：Kini Hemant K; Walton S Patrick^*

来源：FEBS LETTERS, 2007, 581(29): 5611-5616.

DOI：10.1016/j.febslet.2007.11.010

• single-stranded RNA
• siRNA
• Dicer
• divalent cation
• PAZ domain

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 50-phosphate with greater affinity versus those with a 50-hydroxyl. In addition, 30-biotinylated ssRNAs are bound by Dicer with lower affinity than 30-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.

• 出版日期2007-12-11