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.
Gancz
Dana
D
Department of Cell and Developmental Biology, Sackler School of Medicine, Tel Aviv University, Tel Aviv 69978, Israel.
Donin
Natalie
N
Fishelson
Zvi
Z
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