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Disruption of GW bodies impairs mammalian RNA interference

An Erratum to this article was published on 02 December 2005

Abstract

The GW182 RNA-binding protein was initially shown to associate with a specific subset of mRNAs and to reside within discrete cytoplasmic foci named GW bodies (GWBs)1. GWBs are enriched in proteins that are involved in mRNA degradation2. Recent reports have shown that exogenously introduced human Argonaute-2 (Ago2) is also enriched in GWBs, indicating that RNA interference function may be somehow linked to these structures3,4. In this report, we demonstrate that endogenous Ago2 and transfected small interfering RNAs (siRNAs) are also present within these same cytoplasmic bodies and that the GW182 protein interacts with Ago2. Disruption of these cytoplasmic foci in HeLa cells interferes with the silencing capability of a siRNA that is specific to lamin-A/C. Our data support a model in which GW182 and/or the microenvironment of the cytoplasmic GWBs contribute to the RNA-induced silencing complex and to RNA silencing.

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Figure 1: Ago2 associates with GW182 and localizes to GWBs.
Figure 2: Transiently transfected fluorescently labelled siRNAs localize to GWBs.
Figure 3: Disassembly of GWBs correlates with the loss of RNAi activity.
Figure 4: RNA interference is disrupted by sequential transfection of siRNA to GW182 and siRNA to endogenous reporter lamin-A/C.

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Acknowledgements

We thank S. Jablonski for kindly providing the Cy3-labelled luciferase siRNA, T. Hobman for providing the Ago2 rabbit antibody and cDNA, and L.M. Andino for technical help with the RNAse protection assay. This work was supported in part by the Canadian Institutes for Health Research Grant MOP-38034, Canadian Breast Cancer Research Foundation Grant 16992, and the National Institutes of Health Grants AI47859 and AI39645. M.J.F. holds the Arthritis Society Chair at the University of Calgary. A preliminary report in abstract form describing this work was presented at the Tenth Annual Meeting of the RNA Society, Banff, Alberta, Canada, 24–29 May 2005.

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Correspondence to Edward K.L. Chan.

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Supplementary figures S1, S2, S3 and S4 (PDF 710 kb)

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Jakymiw, A., Lian, S., Eystathioy, T. et al. Disruption of GW bodies impairs mammalian RNA interference. Nat Cell Biol 7, 1267–1274 (2005). https://doi.org/10.1038/ncb1334

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