Involvement of the c-jun N-terminal kinases JNK1 and JNK2 in complement-mediated cell death.
Cell death and survival signals activated by the complement membrane attack complex C5b-9 play important roles in complement-associated diseases and in antibody-based cancer therapy. Here, we investigated the involvement of the JNK mitogen-activated protein kinase in C5b-9-induced cell lysis. Necrotic-type cell death regulation by JNK1 and JNK2 was selectively studied in mouse fibroblasts and human K562, HeLa and 293T cells. C5b-9 induced higher JNK activation than C5b-8. Pretreatment with a JNK inhibitor reduced cell sensitivity to complement-mediated lysis. KO cells deficient in either JNK1 or JNK2 were less sensitive to lysis than WT cells. This correlated with lower C3 and C5b-9 deposition on KO cells. Furthermore, silencing of JNK1 or JNK2 expression by RNA interference decreased cell lysis by complement. Reconstitution of JNK2 into JNK2-/- cells and over expression of JNK2 in WT cells increased C3 and C5b-9 deposition as well as cell sensitivity to complement-mediated lysis. Pretreatment of cells with the phosphotyrosine phosphatase inhibitor phenylarsine oxide increased JNK activation and JNK-dependent complement-mediated necrotic death of WT and JNK2-/- KO cells but not of JNK1-/- KO cells. The JNK inhibitor and PAO had no effect on complement-mediated lysis in cells lacking Bid, suggesting involvement of Bid in the JNK lytic pathway. Our results demonstrate that complement C5b-9 induce a JNK/Bid-dependent and JNK-independent necrotic cell death. Both JNK1 and JNK2 have cytotoxic potential, however JNK2 is the primary signal transducer.
Department of Cell and Developmental Biology, Sackler School of Medicine, Tel Aviv University, Tel Aviv 69978, Israel.
Dana
D
Gancz
Natalie
N
Donin
Zvi
Z
Fishelson
D000818:Animals; D001152:Arsenicals / Q000494:pharmacology; D051023:BH3 Interacting Domain Death Agonist Protein / Q000378:metabolism; D016923:Cell Death / Q000276:immunology; D045744:Cell Line, Tumor; D015938:Complement Membrane Attack Complex / Q000276:immunology*; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D020868:Gene Silencing / Q000187:drug effects; D006801:Humans; D051379:Mice; D048055:Mitogen-Activated Protein Kinase 8 / Q000172:deficiency / Q000378:metabolism*; D048056:Mitogen-Activated Protein Kinase 9 / Q000172:deficiency / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D001152:Arsenicals; D051023:BH3 Interacting Domain Death Agonist Protein; D015938:Complement Membrane Attack Complex; D034741:RNA, Small Interfering; C029341:oxophenylarsine; D014443:Tyrosine; D048056:Mitogen-Activated Protein Kinase 9; D048055:Mitogen-Activated Protein Kinase 8; D010749:Phosphoprotein Phosphatases