Abstract
RISC, the RNA-induced silencing complex, uses short interfering RNAs (siRNAs) or micro RNAs (miRNAs) to select its targets in a sequence-dependent manner. Key RISC components are Argonaute proteins, which contain two characteristic domains, PAZ and PIWI. PAZ is highly conserved and is found only in Argonaute proteins and Dicer. We have solved the crystal structure of the PAZ domain of Drosophila Argonaute2. The PAZ domain contains a variant of the OB fold, a module that often binds single-stranded nucleic acids. PAZ domains show low-affinity nucleic acid binding, probably interacting with the 3′ ends of single-stranded regions of RNA. PAZ can bind the characteristic two-base 3′ overhangs of siRNAs, indicating that although PAZ may not be a primary nucleic acid binding site in Dicer or RISC, it may contribute to the specific and productive incorporation of siRNAs and miRNAs into the RNAi pathway.
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Acknowledgements
We thank E. Enemark and M. Carmell for help with refinement and figures, A. Caudy for aid in characterizing the biochemical properties of GST-PAZ, M. Myers for mass spectrometry and A. Heroux (beamline X26C) for support with data collection at the National Synchrotron Light Source (NSLS) at Brookhaven National Laboratory. The NSLS is supported by the US Department of Energy, Division of Materials Sciences and Division of Chemical Sciences. J.J.S. is a Bristol-Myers Squibb Predoctoral Fellow, N.H.T. is a Leslie C. Quick Jr. Predoctoral Fellow. This work was supported by the Watson School of Biological Sciences (L.J.) and the US National Institutes of Health (G.J.H.). G.J.H. is a Rita Allen Foundation Scholar and is supported by an Innovator Award from the US Army Breast Cancer Research Fund.
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Song, JJ., Liu, J., Tolia, N. et al. The crystal structure of the Argonaute2 PAZ domain reveals an RNA binding motif in RNAi effector complexes. Nat Struct Mol Biol 10, 1026–1032 (2003). https://doi.org/10.1038/nsb1016
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DOI: https://doi.org/10.1038/nsb1016
