Global Research Laboratory, School of Biological Science/Bio-Max Institute, Seoul National University, Korea.
Cell Death Differ. 2012 Jul;19(7):1196-207. doi: 10.1038/cdd.2011.209. Epub 2012 Jan 13.
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors, TRAIL-R1 (DR4) and TRAIL-R2 (DR5), promote the selective clearing of various malignancies by inducing apoptosis, holding the promise as a potent therapeutic agent for anticancer. Though DR4 and DR5 have high sequence similarity, differential regulation of both receptors in human tumor cells remains largely unexplored. Here, we repot that golgi-specific Asp-His-His-Cys (DHHC) zinc finger protein (GODZ) regulates TRAIL/DR4-mediated apoptosis. Using the SOS protein recruitment-yeast two-hybrid screening, we isolated GODZ that interacted with the death domain of DR4. GODZ binds to DR4, but not to DR5, through the DHHC and the C-terminal transmembrane domain. Expression level of GODZ affects apoptosis of tumor cells triggered by TRAIL, but not that induced by TNF-α/cycloheximide (CHX) or DNA-damaging drugs. In parallel, GODZ functions to localize DR4 to the plasma membrane (PM) via DHHC motif. Also, introduction of mutation into the cysteine-rich motif of DR4 results in its mistargeting and attenuates TRAIL- or GODZ-mediated apoptosis. Interestingly, GODZ expression is highly downregulated in Hep-3B tumor cells, which show resistance to TRAIL. However, reconstitution of GODZ expression enhances the targeting of DR4 to cell surface and sensitizes Hep-3B cells to TRAIL. Taken together, these data establish that GODZ is a novel DR4-selective regulator responsible for targeting of DR4 to the PM, and thereby for TRAIL-induced apoptosis.
肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其受体 TRAIL-R1(DR4)和 TRAIL-R2(DR5)通过诱导细胞凋亡促进各种恶性肿瘤的选择性清除,有望成为一种有效的抗癌治疗药物。虽然 DR4 和 DR5 具有高度的序列相似性,但人类肿瘤细胞中这两种受体的差异调节在很大程度上仍未得到探索。在这里,我们报告高尔基特异性天冬氨酸-组氨酸-组氨酸-半胱氨酸(DHHC)锌指蛋白(GODZ)调节 TRAIL/DR4 介导的细胞凋亡。我们利用 SOS 蛋白募集-酵母双杂交筛选,分离出与 DR4 死亡结构域相互作用的 GODZ。GODZ 通过 DHHC 和 C 末端跨膜结构域与 DR4 结合,但不与 DR5 结合。GODZ 的表达水平影响 TRAIL 触发的肿瘤细胞凋亡,但不影响 TNF-α/环己酰亚胺(CHX)或 DNA 损伤药物诱导的凋亡。同时,GODZ 通过 DHHC 基序将 DR4 定位于质膜(PM)。此外,将突变引入 DR4 的富含半胱氨酸的基序会导致其靶向错误,并减弱 TRAIL 或 GODZ 介导的细胞凋亡。有趣的是,GODZ 在对 TRAIL 具有抗性的 Hep-3B 肿瘤细胞中高度下调表达。然而,GODZ 表达的重建增强了 DR4 向细胞表面的靶向,并使 Hep-3B 细胞对 TRAIL 敏感。总之,这些数据表明 GODZ 是一种新的 DR4 选择性调节剂,负责将 DR4 靶向到 PM,从而促进 TRAIL 诱导的细胞凋亡。