TY - CHAP
T1 - Analysis of cytokine- and influenza A virus-driven RIPK3 necrosome formation
AU - Thapa, Roshan J.
AU - Nogusa, Shoko
AU - Balachandran, Siddharth
N1 - Publisher Copyright:
© 2018, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2018
Y1 - 2018
N2 - 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).
AB - 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).
KW - Animals
KW - Cells, Cultured
KW - Cytokines/pharmacology
KW - Embryo, Mammalian/drug effects
KW - Fibroblasts/drug effects
KW - Influenza A virus/drug effects
KW - Mice
KW - Necrosis
KW - Orthomyxoviridae Infections/drug therapy
KW - Phosphorylation
KW - Receptor-Interacting Protein Serine-Threonine Kinases/metabolism
KW - Signal Transduction
UR - http://www.scopus.com/inward/record.url?scp=85052406121&partnerID=8YFLogxK
UR - https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=purepublist2023&SrcAuth=WosAPI&KeyUT=WOS:000459305800010&DestLinkType=FullRecord&DestApp=WOS
U2 - 10.1007/978-1-4939-8754-2_9
DO - 10.1007/978-1-4939-8754-2_9
M3 - Chapter
C2 - 30136233
VL - 1857
T3 - Methods in molecular biology (Clifton, N.J.)
SP - 93
EP - 99
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -