Abstract
Mixed lineage kinase domain-like protein (Mlkl) was recently found to interact with receptor interacting protein 3 (Rip3) and to be essential for tumor necrosis factor (TNF)-induced programmed necrosis (necroptosis) in cultured cell lines. We have generated Mlkl-deficient mice by transcription activator-like effector nucleases (TALENs)-mediated gene disruption and found Mlkl to be dispensable for normal mouse development as well as immune cell development. Mlkl-deficient mouse embryonic fibroblasts (MEFs) and macrophages both showed resistance to necrotic but not apoptotic stimuli. Mlkl-deficient MEFs and macrophages were indistinguishable from wild-type cells in their ability to activate NF-κB, ERK, JNK, and p38 in response to TNF and lipopolysaccharides (LPS), respectively. Consistently, Mlkl-deficient macrophages and mice exhibited normal interleukin-1β (IL-1β), IL-6, and TNF production after LPS treatment. Mlkl deficiency protects mice from cerulean-induced acute pancreatitis, a necrosis-related disease, but has no effect on polymicrobial septic shock-induced animal death. Our results provide genetic evidence for the role of Mlkl in necroptosis.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Amino Acid Chloromethyl Ketones / pharmacology
-
Animals
-
Apoptosis*
-
Base Sequence
-
Cell Line
-
Interleukin-1beta / metabolism
-
Interleukin-6 / metabolism
-
Lipopolysaccharides / toxicity
-
Macrophages / drug effects
-
Macrophages / metabolism
-
Male
-
Mice
-
Mice, Inbred C57BL
-
Mice, Knockout
-
Mitochondria / drug effects
-
Mitochondria / metabolism
-
Mitogen-Activated Protein Kinase Kinases / metabolism
-
NF-kappa B / metabolism
-
Necrosis
-
Protein Kinases / deficiency
-
Protein Kinases / genetics
-
Protein Kinases / metabolism*
-
Receptor-Interacting Protein Serine-Threonine Kinases / deficiency
-
Receptor-Interacting Protein Serine-Threonine Kinases / genetics
-
Receptor-Interacting Protein Serine-Threonine Kinases / metabolism
-
Signal Transduction / drug effects
-
Tumor Necrosis Factor-alpha / pharmacology
-
Tumor Necrosis Factors / metabolism
Substances
-
Amino Acid Chloromethyl Ketones
-
Interleukin-1beta
-
Interleukin-6
-
Lipopolysaccharides
-
NF-kappa B
-
Tumor Necrosis Factor-alpha
-
Tumor Necrosis Factors
-
benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
-
MLKL protein, mouse
-
Protein Kinases
-
Receptor-Interacting Protein Serine-Threonine Kinases
-
Ripk3 protein, mouse
-
Mitogen-Activated Protein Kinase Kinases