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Immunoblot Analysis of the Regulation of TNF Receptor Family-Induced NF-κB Signaling by c-IAP Proteins

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NF-κB Transcription Factors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2366))

Abstract

Proper maintenance of organismal homeostasis, development, and immune defense requires precise regulation of survival and signaling pathways. Inhibitor of apoptosis (IAP) proteins are evolutionarily conserved regulators of cell death and immune signaling that impact numerous cellular processes. Although initially characterized as inhibitors of apoptosis, the ubiquitin ligase activity of IAP proteins is critical for modulating various signaling pathways (e.g., NF-κB, MAPK) and cell survival. Cellular IAP1 and 2 regulate the pro-survival canonical NF-κB pathway by ubiquitinating RIP1 and themselves thus enabling recruitment of kinase (IKK) and E3 ligase (LUBAC) complexes. On the other hand, c-IAP1 and c-IAP2 are negative regulators of noncanonical NF-κB signaling by promoting ubiquitination and consequent proteasomal degradation of the NF-κB-inducing kinase NIK. Here we describe the involvement of c-IAP1 and c-IAP2 in NF-κB signaling and provide detailed methodology for examining functional roles of c-IAPs in these pathways.

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Acknowledgments

We thank researchers at Genentech who helped with suggestions, reagents, and comments. The authors are employees of Genentech.

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Authors and Affiliations

  1. Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA, USA

    Eugene Varfolomeev, Tatiana Goncharov & Domagoj Vucic

Authors
  1. Eugene Varfolomeev
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  2. Tatiana Goncharov
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  3. Domagoj Vucic
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Corresponding author

Correspondence to Domagoj Vucic .

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Editors and Affiliations

  1. Centre for Molecular Immunology and Inflammation, Department of Immunology and Inflammation, Imperial College London, London, UK

    Guido Franzoso

  2. Department of Biotechnological and Applied Clinical Sciences, University of L’Aquila, L’Aquila, Italy

    Francesca Zazzeroni

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Varfolomeev, E., Goncharov, T., Vucic, D. (2021). Immunoblot Analysis of the Regulation of TNF Receptor Family-Induced NF-κB Signaling by c-IAP Proteins. In: Franzoso, G., Zazzeroni, F. (eds) NF-κB Transcription Factors. Methods in Molecular Biology, vol 2366. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1669-7_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1669-7_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1668-0

  • Online ISBN: 978-1-0716-1669-7

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