title: CD59 signaling and membrane pores drive Syk-dependent erythrocyte necroptosis.
abstract: Mature erythrocytes (red blood cells (RBCs)) undergo the programmed cell death (PCD) pathway of necroptosis in response to bacterial pore-forming toxins (PFTs) that target human CD59 (hCD59) but not hCD59-independent PFTs. Here, we investigate the biochemical mechanism of RBC necroptosis with a focus on the mechanism of induction and the minimal requirements for such RBC death. Binding or crosslinking of the hCD59 receptor led to Syk-dependent induction of vesiculated morphology (echinocytes) that was associated with phosphorylation of Band 3 and was required for Fas ligand (FasL) release. FasL-dependent phosphorylation of receptor-interacting protein kinase 1 (RIP1) in combination with plasma membrane pore formation was required for execution of RBC necroptosis. RIP1 phosphorylation led to the phosphorylation of RIP3, which was also critical for RBC necroptosis. Notably, RBC necroptosis was mediated by FasL and not by other candidate inducers, including tumor necrosis factor alpha (TNF-α) and TNF-related apoptosis-inducing ligand (TRAIL). Other types of RBC damage, such as eryptotic damage, failed to induce necroptosis when combined with hCD59 crosslinking. This work sheds light on the requirements for this recently discovered PCD in RBCs and provides a clear picture of the biochemical mechanism of induction of RBC necroptosis.
authors.affiliation: Department of Pediatrics, Columbia University, New York, NY, USA.
authors.forename: T J
authors.initials: TJ
authors.lastname: LaRocca
authors.affiliation: Institute of Human Nutrition, Columbia University, New York, NY, USA.
authors.forename: E A
authors.initials: EA
authors.lastname: Stivison
authors.affiliation: Department of Pediatrics, Columbia University, New York, NY, USA.
authors.forename: T
authors.initials: T
authors.lastname: Mal-Sarkar
authors.affiliation: Department of Pediatrics, Columbia University, New York, NY, USA.
authors.forename: T A
authors.initials: TA
authors.lastname: Hooven
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, USA.
authors.forename: E A
authors.initials: EA
authors.lastname: Hod
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, USA.
authors.forename: S L
authors.initials: SL
authors.lastname: Spitalnik
authors.affiliation: Department of Pediatrics, Columbia University, New York, NY, USA.
authors.forename: A J
authors.initials: AJ
authors.lastname: Ratner
mesh_terms: D018958:CD55 Antigens / Q000276:immunology / Q000378:metabolism; D018957:CD59 Antigens / Q000276:immunology / Q000378:metabolism*; D002462:Cell Membrane / Q000473:pathology; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004912:Erythrocytes / Q000473:pathology*; D053222:Fas Ligand Protein / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D009336:Necrosis / Q000276:immunology*; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D010766:Phosphorylation; D052899:Pore Forming Cytotoxic Proteins / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000378:metabolism; D015398:Signal Transduction; D000072377:Syk Kinase; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: C119724:AGFG1 protein, human; D018958:CD55 Antigens; D018957:CD59 Antigens; D003432:Cross-Linking Reagents; C505661:FASLG protein, human; D053222:Fas Ligand Protein; D047908:Intracellular Signaling Peptides and Proteins; D028861:Nuclear Pore Complex Proteins; D052899:Pore Forming Cytotoxic Proteins; D016601:RNA-Binding Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D014409:Tumor Necrosis Factor-alpha; C051590:CD59 protein, human; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C119705:RIPK3 protein, human; D053422:Receptor-Interacting Protein Serine-Threonine Kinases