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The mitochondrial Nod-like receptor NLRX1 modifies apoptosis through SARM1

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Abstract

NLRX1, the mitochondrial NOD-like receptor (NLR), modulates apoptosis in response to both intrinsic and extrinsic cues. Insights into the mechanism of how NLRX1 influences apoptosis remain to be determined. Here, we demonstrate that NLRX1 associates with SARM1, a protein with a toll/interleukin-1 receptor (TIR)-containing domain also found in adaptor proteins downstream of toll-like receptors, such as MyD88. While a direct role of SARM1 in innate immunity is unclear, the protein plays essential roles in Wallerian degeneration (WD), a type of neuronal catabolism occurring following axonal severing or damage. In non-neuronal cells, we found that endogenous SARM1 was equally distributed in the cytosol and the mitochondrial matrix, where association with NLRX1 occurred. In these cells, the apoptotic role of NLRX1 was fully dependent on SARM1, indicating that SARM1 was downstream of NLRX1 in apoptosis regulation. In primary murine neurons, however, Wallerian degeneration induced by vinblastine or NGF deprivation occurred in SARM1- yet NLRX1-independent manner, suggesting that WD requires the cytosolic pool of SARM1 or that NLRX1 levels in neurons are too low to contribute to WD regulation. Together, these results shed new light into the mechanisms through which NLRX1 controls apoptosis and provides evidence of a new link between NLR and TIR-containing proteins.

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Acknowledgements

This project was supported by grants from Canadian Institutes of Health Research (CIHR) and Natural Sciences and Engineering Research Council of Canada (NSERC) to SEG and the CIHR Vanier Canada Graduate Scholarship to SAK.

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Authors and Affiliations

  1. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, M5S 1A8, Canada

    Samuel A. Killackey, Fraser Soares, Ashley B. Zhang, Mena Abdel-Nour & Stephen E. Girardin

  2. Department of Immunology, University of Toronto, Toronto, ON, M5S 1A8, Canada

    Muhammed A. Rahman, Dana J. Philpott & Stephen E. Girardin

Authors
  1. Samuel A. Killackey
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  2. Muhammed A. Rahman
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  3. Fraser Soares
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  4. Ashley B. Zhang
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  5. Mena Abdel-Nour
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  6. Dana J. Philpott
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  7. Stephen E. Girardin
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Correspondence to Stephen E. Girardin.

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Online Resource Fig. 1

Anti-SARM1 antibody labels numerous non-specific bands. a, Immunoblot of whole brain extracts taken from P0-4 pups of WT and Sarm1−/−. b, Immunoblot of cytosolic and mitochondrial fractions from WT or SARM1−/− CRISPR cells. Blots were stained with either antibodies from GeneTex (left) or Cell Signaling Technology (right) (PPTX 377 KB)

Online Resource Fig. 2

Validation of cell death induction following apoptotic treatments. TNF-α (10 ng/mL) + CHX (10 µg/mL) for 6 h or A23187 (0.625, 1.25, 2.5 µM) for 20 h (PPTX 108 KB)

Online Resource Fig. 3

Validation of SARM1 knockdown using shRNA. Arrow designates SARM1 band at approximately 73 kDa (PPTX 80 KB)

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Killackey, S.A., Rahman, M.A., Soares, F. et al. The mitochondrial Nod-like receptor NLRX1 modifies apoptosis through SARM1. Mol Cell Biochem 453, 187–196 (2019). https://doi.org/10.1007/s11010-018-3444-3

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  • DOI: https://doi.org/10.1007/s11010-018-3444-3

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