In vitro binding of single-stranded RNA by human Dicer

Hemant K. Kini, S. Patrick Walton

    Research output: Contribution to journalArticle

    • 15 Citations

    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.

    Original languageEnglish (US)
    Pages (from-to)5611-5616
    Number of pages6
    JournalFEBS Letters
    Volume581
    Issue number29
    DOIs
    StatePublished - Dec 11 2007

    Profile

    RNA
    In Vitro Techniques
    Hydroxyl Radical
    Double-Stranded RNA
    Divalent Cations
    Small Interfering RNA
    Phosphates
    Bibliography of Medicine

    Keywords

    • Dicer
    • Divalent cation
    • PAZ domain
    • Single-stranded RNA
    • siRNA

    ASJC Scopus subject areas

    • Biochemistry
    • Biophysics
    • Molecular Biology

    Cite this

    In vitro binding of single-stranded RNA by human Dicer. / Kini, Hemant K.; Walton, S. Patrick.

    In: FEBS Letters, Vol. 581, No. 29, 11.12.2007, p. 5611-5616.

    Research output: Contribution to journalArticle

    Kini, Hemant K.; Walton, S. Patrick / In vitro binding of single-stranded RNA by human Dicer.

    In: FEBS Letters, Vol. 581, No. 29, 11.12.2007, p. 5611-5616.

    Research output: Contribution to journalArticle

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