Recognition of siRNA asymmetry by TAR RNA binding protein

Joseph A. Gredell, Michael J. Dittmer, Ming Wu, Christina Chan, S. Patrick Walton

    Research output: Research - peer-reviewArticle

    • 32 Citations

    Abstract

    The recognition of small interfering RNAs (siRNAs) by the RNA-induced silencing complex (RISC) and its precursor, the RISC loading complex (RLC), is a key step in the RNA interference pathway that controls the subsequent sequence-specific mRNA degradation. In Drosophila, selection of the guide strand has been shown to be mediated by RLC protein R2D2, which senses the relative hybridization stability between the two ends of the siRNA. A protein with similar function has yet to be conclusively identified in humans. We show here that human TAR RNA binding protein (TRBP) alone can bind siRNAs in vitro and sense their asymmetry. We also show that TRBP can bind 21-nucleotide single-stranded RNAs, though with far lower affinity than for double-stranded siRNA, and that TRBP cross-links preferentially to the 3'-ends of the guide strands of siRNAs. This suggests that TRBP binding depends both on the sequences of the siRNA strands and on the relative hybridization stability of the ends of the duplex. Together, these results demonstrate the importance of the siRNA-TRBP interaction in the selection of the siRNA guide strand in RNAi.

    LanguageEnglish (US)
    Pages3148-3155
    Number of pages8
    JournalBiochemistry
    Volume49
    Issue number14
    DOIs
    StatePublished - Apr 13 2010

    Profile

    Small Interfering RNA
    trans-activation responsive RNA-binding protein
    RNA-Induced Silencing Complex
    RNA Interference
    Proteins
    RNA
    RNA Stability
    Protein Binding
    Drosophila
    Nucleotides
    In Vitro Techniques
    Reduced instruction set computing
    Degradation
    Messenger RNA

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Recognition of siRNA asymmetry by TAR RNA binding protein. / Gredell, Joseph A.; Dittmer, Michael J.; Wu, Ming; Chan, Christina; Walton, S. Patrick.

    In: Biochemistry, Vol. 49, No. 14, 13.04.2010, p. 3148-3155.

    Research output: Research - peer-reviewArticle

    Gredell, JA, Dittmer, MJ, Wu, M, Chan, C & Walton, SP 2010, 'Recognition of siRNA asymmetry by TAR RNA binding protein' Biochemistry, vol 49, no. 14, pp. 3148-3155. DOI: 10.1021/bi902189s
    Gredell JA, Dittmer MJ, Wu M, Chan C, Walton SP. Recognition of siRNA asymmetry by TAR RNA binding protein. Biochemistry. 2010 Apr 13;49(14):3148-3155. Available from, DOI: 10.1021/bi902189s
    Gredell, Joseph A. ; Dittmer, Michael J. ; Wu, Ming ; Chan, Christina ; Walton, S. Patrick. / Recognition of siRNA asymmetry by TAR RNA binding protein. In: Biochemistry. 2010 ; Vol. 49, No. 14. pp. 3148-3155
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