Analysis of cytokine- and influenza A virus-driven RIPK3 necrosome formation

Roshan J. Thapa, Shoko Nogusa, Siddharth Balachandran

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

4 Scopus citations

Abstract

In multicellular organisms, regulated cell death plays a vital role in development, adult tissue homeostasis, and clearance of damaged or infected cells. Necroptosis is one such form of regulated cell death, characterized by its reliance on receptor-interacting protein kinase 3 (RIPK3). Once activated, RIPK3 nucleates a protein complex, termed the “necrosome,” which includes the adaptors RIPK1 and FADD, and the effector protein MLKL. From the necrosome, RIPK3 phosphorylates MLKL to drive necroptosis, and can also induce RIPK1/FADD-mediated apoptosis, via caspase-8. Assembly of the necrosome thus serves as a useful readout of RIPK3 activation. In this chapter, we describe molecular methods for examining necrosome activation in response to the cytokines TNF-α, IFN-β, and IFN-γ, and upon infection with influenza A virus (IAV).

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages93-99
Number of pages7
Volume1857
DOIs
StatePublished - 2018

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press Inc.
ISSN (Print)1064-3745

Keywords

  • Animals
  • Cells, Cultured
  • Cytokines/pharmacology
  • Embryo, Mammalian/drug effects
  • Fibroblasts/drug effects
  • Influenza A virus/drug effects
  • Mice
  • Necrosis
  • Orthomyxoviridae Infections/drug therapy
  • Phosphorylation
  • Receptor-Interacting Protein Serine-Threonine Kinases/metabolism
  • Signal Transduction

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