FADD is required for DR4-and DR5-mediated apoptosis - Lack of trail-induced apoptosis in FADD-deficient mouse embryonic fibroblasts
FADD is required for DR4-and DR5-mediated apoptosis - Lack of trail-induced apoptosis in FADD-deficient mouse embryonic fibroblasts
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DOI：
10.1074/jbc.c000284200
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发表时间：
2000-08-18
影响因子：
4.8
通讯作者：
Winoto, A
中科院分区：
文献类型：
--
作者：
Kuang, AA;Diehl, GE;Winoto, A
肿瘤坏死因子相关凋亡诱导配体（tumor necrosis factor-related apoptosis-inducing ligand，TRAIL）是肿瘤坏死因子家族的一员，能杀伤多种肿瘤细胞，但不能杀伤正常细胞。TRAIL诱导的人类细胞凋亡由其受体DR 4（TRAIL-R1）和DR 5（TRAIL-R2）介导。然而，这些受体下游的信号分子是由什么构成的，仍然存在很大的争议。使用FADD显性负性分子，几个研究小组就FADD在TRAIL诱导的细胞凋亡中的作用得出了不同的结论。最近，使用FADD缺陷（-/-）小鼠胚胎成纤维细胞，Yeh等（Yeh，W. C.的方法，Pompa，J.L.，McCurrach，M. E、Shu，H.- B.，Elia，A. J，，Shahinian，A，Ng，M.，Wakeham，A，Khoo，W.，米切尔，K.，El-Deiry，W.美国，Lowe，S. W.，Goeddel，D.五、和Mak，T，W.（1998）Science 279，1954-1958）得出结论，DR 4利用FADD-非依赖性凋亡途径。然而，后一个实验涉及瞬时过表达，这通常导致含有死亡结构域的受体的非特异性聚集。为了在更生理的环境中解决这个问题，我们将小鼠DR 4/5、人DR 4或人DR 5稳定转染到FADD（-/-）小鼠胚胎成纤维细胞中。我们发现用TRAIL受体稳定转染的FADD（-/-）MEF细胞对TRAIL介导的细胞死亡具有抗性。相反，稳定转染到杂合FADD（+/-）细胞或用FADD逆转录病毒构建体重建的FADD（-/-）细胞中的TRAIL受体对TRAIL细胞毒性效应敏感。我们得出结论，FADD是需要DR 4和DR 5介导的细胞凋亡。
TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is a member of the tumor necrosis factor family that can kill a wide variety of tumor cells but not normal cells. TRAIL-induced apoptosis in humans is mediated by its receptors DR4 (TRAIL-R1) and DR5 (TRAIL-R2). What constitutes the signaling molecules downstream of these receptors, however, remains highly controversial. Using the FADD dominant negative molecule, several groups have reached different conclusions with respect to the role of FADD in TRAIL-induced apoptosis. More recently, using FADD-deficient (-/-) mouse embryonic fibroblasts, Yeh ct at (Yeh, W. C., Pompa, J. L., McCurrach, M. E., Shu, H.-B., Elia, A. J,, Shahinian, A, Ng, M., Wakeham, A, Khoo, W., Mitchell, K., El-Deiry, W. S., Lowe, S. W., Goeddel, D. V., and Mak, T,W. (1998) Science 279, 1954-1958) concluded that DR4 utilizes a FADD-independent apoptotic pathway. The latter experiment, however, involved transient overexpression, which often leads to nonspecific aggregation of death domain-containing receptors. To address this issue in a more physiological setting, we stably transfected mouse DR4/5, human DR4, or human DR5 into FADD(-/-) mouse embryonic fibroblast cells. We showed that FADD(-/-) MEF cells stably transfected with TRAIL receptors are resistant to TRAIL-mediated cell death. In contrast, TRAIL receptors stably transfected into heterozygous FADD(+/-) cells or FADD(-/-) cells reconstituted with a FADD retroviral construct are sensitive to the TRAIL cytotoxic effect. We conclude that FADD is required for DR4- and DR5-mediated apoptosis.