TRIM23 mediates virus-induced autophagy via activation of TBK1

Abstract

Autophagy and interferon (IFN)-mediated innate immunity are critical antiviral defence mechanisms, and recent evidence indicated that tripartite motif (TRIM) proteins are important regulators of both processes. Although the role of TRIM proteins in modulating antiviral cytokine responses has been well established, much less is known about their involvement in autophagy in response to different viral pathogens. Through a targeted RNAi screen examining the relevance of selected TRIM proteins in autophagy induced by herpes simplex virus 1 (HSV-1), encephalomyocarditis virus (EMCV) and influenza A virus (IAV), we identified several TRIM proteins that regulate autophagy in a virus-species-specific manner, as well as a few TRIM proteins that were essential for autophagy triggered by all three viruses and rapamycin, among them TRIM23. TRIM23 was critical for autophagy-mediated restriction of multiple viruses, and this activity was dependent on both its RING E3 ligase and ADP-ribosylation factor (ARF) GTPase activity. Mechanistic studies revealed that unconventional K27-linked auto-ubiquitination of the ARF domain is essential for the GTP hydrolysis activity of TRIM23 and activation of TANK-binding kinase 1 (TBK1) by facilitating its dimerization and ability to phosphorylate the selective autophagy receptor p62. Our work identifies the TRIM23-TBK1-p62 axis as a key component of selective autophagy and further reveals a role for K27-linked ubiquitination in GTPase-dependent TBK1 activation.

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Fig. 1: TRIM proteins modulate viral-induced autophagy in a virus species-specific manner.
Fig. 2: TRIM23 is essential for virus-induced autophagy.
Fig. 3: K27-linked auto-ubiquitination of the ARF domain of TRIM23 is necessary for its autophagy function.
Fig. 4: ARF ubiquitination is required for the GTP hydrolysis activity of TRIM23 and its localization to autophagosomes.
Fig. 5: TRIM23 interacts with TBK1 and p62.
Fig. 6: TRIM23 GTPase activates TBK1 to phosphorylate p62.

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Acknowledgements

The authors thank A. García-Sastre (Icahn School of Medicine at Mount Sinai) and J. Jung (University of Southern California) for the TRIM cDNA library, and S. Rabkin (Harvard) for providing mutant HSV-1. The authors also thank M. Ericsson (Harvard Electron Microscopy Facility) for assistance with sample preparation and S. Hwang (The University of Chicago) for discussions. This study was supported in part by the US National Institutes of Health grants R01 AI087846 and R21 AI118509 (to M.U.G.) and R01 GM112508 (to O.P.). K.M.J.S. and F.F. were supported by fellowships from the German Research Foundation (SP 1600/1-1 and FU 949/1-1, respectively). G.P.-R., J.K., J.M. and M.V. were supported by the Intramural Research Program of the NIH (National Heart, Lung, and Blood Institute). M.A.Z. received support by NIH training grant T32 GM007183.

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K.M.J.S., S.G. and M.U.G. conceived and designed the experiments. K.M.J.S. and S.G. performed and analysed all experiments, except those in Supplementary Fig. 4f (M.A.Z.), Supplementary Fig. 2b,c (F.F.) and Fig. 5e (Z.M.P.). G.J.B. performed mutagenesis experiments. J.K., G.P.-R., J.M. and M.V. provided TRIM23 −/− and WT MEFs. C.L. contributed reagents, materials and analysis tools for experiments. O.P. performed the TRIM23 ARF structure modelling. K.M.J.S. and M.U.G. wrote the manuscript.

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Correspondence to Michaela U. Gack.

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Sparrer, K.M.J., Gableske, S., Zurenski, M.A. et al. TRIM23 mediates virus-induced autophagy via activation of TBK1. Nat Microbiol 2, 1543–1557 (2017). https://doi.org/10.1038/s41564-017-0017-2

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