@article{54b2998e640b4ff59db15abaac96f8c6,
title = "Structure and conserved RNA binding of the PAZ domain",
abstract = "The discovery of RNA-mediated gene-silencing pathways, including RNA interference, highlights a fundamental role of short RNAs in eukaryotic gene regulation and antiviral defence. Members of the Dicer and Argonaute protein families are essential components of these RNA-silencing pathways. Notably, these two families possess an evolutionarily conserved PAZ (Piwi/Argonaute/Zwille) domain whose biochemical function is unknown. Here we report the nuclear magnetic resonance solution structure of the PAZ domain from Drosophila melanogaster Argonaute 1 (Ago1). The structure consists of a left-handed, six-stranded β-barrel capped at one end by two α-helices and wrapped on one side by a distinctive appendage, which comprises a long β-hairpin and a short α-helix. Using structural and biochemical analyses, we demonstrate that the PAZ domain binds a 5-nucleotide RNA with 1:1 stoichiometry. We map the RNA-binding surface to the open face of the β-barrel, which contains amino acids conserved within the PAZ domain family, and we define the 5′-to-3′ orientation of single-stranded RNA bound within that site. Furthermore, we show that PAZ domains from different human Argonaute proteins also bind RNA, establishing a conserved function for this domain.",
author = "Yan, {Kelley S.} and Sherry Yan and Amjad Farooq and Arnold Han and Lei Zeng and Zhou, {Ming Ming}",
note = "Funding Information: Acknowledgements We thank A. Ladurner, E. Conti and R. Russell for discussions; G. Stier and S. B{\"a}ckstr{\"o}m for the pETM60 vector; M. Rode for technical support; F. Ciccarreli and P. Bork for help with sequence alignments; and the NMR centre in Frankfurt, Germany for NMR measurement time. This study was supported by the European Molecular Biology Organization (EMBO), the German Research Foundation (DFG), and the Human Frontier Science Program Organization. Funding Information: Acknowledgements We are grateful to G. Hollopeter for generation of the adult rat trigeminal cDNA library and to B. Trueb for providing us with human ANKTM1 cDNA. This work was supported by grants from the Segerfalk Foundation and the Swedish Research Council (P.Z. and E.H.), the American Heart Association (H.C.) and the National Institutes of Health (I.M., D.B. and D.J.). Funding Information: Acknowledgements We thank R. W. Williams for providing a D. melanogaster Ago1 expressed sequenced tag clone, T. Tuschl for complementary DNAs encoding human Argonaute proteins, and T. A. Edwards for discussions. K.S.Y. is a recipient of a National Institutes of Health (NIH) predoctoral training grant fellowship. M.-M.Z. is supported by NIH grants, and is a member of the New York Structural Biology Center.",
year = "2003",
month = nov,
day = "27",
doi = "10.1038/nature02129",
language = "English (US)",
volume = "426",
pages = "469--474",
journal = "Nature",
issn = "0028-0836",
publisher = "Nature Publishing Group",
number = "6965",
}