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Allosteric regulation of Argonaute proteins by miRNAs

Nature Structural & Molecular Biology volume 17, pages 144–150 (2010)Cite this article

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Abstract

Small interfering RNAs (siRNAs) and microRNAs (miRNAs) bind to Argonaute (AGO) family proteins to form a related set of effector complexes that have diverse roles in post-transcriptional gene regulation throughout the eukaryotic lineage. Here sequence and structural analysis of the MID domain of the AGO proteins identified similarities with a family of allosterically regulated bacterial ligand-binding domains. We used in vitro and in vivo approaches to show that certain AGO proteins (those involved in translational repression) have conserved this functional allostery between two distinct sites, one involved in binding miRNA–target duplex and the other in binding the 5′ cap feature (m7GpppG) of eukaryotic mRNAs. This allostery provides an explanation for how miRNA-bound effector complexes may avoid indiscriminate repressive action (mediated through binding interactions with the cap) before full target recognition.

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Figure 1: Bioinformatic analysis of AGO proteins.
Figure 2: Binding of AGO proteins to m7GTP-Sepharose reveals allosteric behavior.
Figure 3: Filter binding assays indicate two allosterically regulated nucleotide binding sites.
Figure 4: Mutational analysis of AGO proteins in Drosophila S2 cells.
Figure 5: Cartoon describing potential role of AGO allostery in promoting translational repression.

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Acknowledgements

We thank S. Dorner for early contributions to the project, E. Izaurralde (Max Planck Institute) for providing the luciferase reporter constructs, H. Zaher (Johns Hopkins Univ. School of Medicine) for mRNA constructs used in filter binding assays, and J. Mendell, G. Seydoux, J. Lorsch, L. Cochella and H. Zaher for helpful comments on the manuscript. J.K.H. was supported by The Samsung Foundation of Culture. The project was supported by funding from the Howard Hughes Medical Institute.

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  1. Sergej Djuranovic and Michelle Kim Zinchenko: These authors contributed equally to this work.

Authors and Affiliations

  1. Howard Hughes Medical Institute and Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Sergej Djuranovic, Michelle Kim Zinchenko, Junho K Hur, Ali Nahvi, Julie L Brunelle, Elizabeth J Rogers & Rachel Green

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Contributions

S.D. performed the bioinformatic analyses and in vitro studies; M.K.Z. performed the in vivo studies; J.K.H., A.N., J.L.B. and E.J.R. performed biochemistry in support of the main experiments; R.G. advised on the project; S.D., M.K.Z. and R.G. prepared the manuscript.

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Correspondence to Rachel Green.

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Djuranovic, S., Zinchenko, M., Hur, J. et al. Allosteric regulation of Argonaute proteins by miRNAs. Nat Struct Mol Biol 17, 144–150 (2010). https://doi.org/10.1038/nsmb.1736

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