JNK1 physically interacts with WW domain-containing oxidoreductase (WOX1) and inhibits WOX1-mediated apoptosis
JNK1 physically interacts with WW domain-containing oxidoreductase (WOX1) and inhibits WOX1-mediated apoptosis
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DOI：
10.1074/jbc.m208373200
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发表时间：
2003-03-14
影响因子：
4.8
通讯作者：
Ensign, A
中科院分区：
文献类型：
--
作者：
Chang, NS;Doherty, J;Ensign, A
c-Jun n -末端激酶（JNK）的短暂激活促进细胞存活，而JNK的持续激活诱导细胞凋亡。牛睾丸透明质酸酶PH-20激活JNK1并保护L929成纤维细胞免受星罗孢素介导的细胞死亡。PH-20还诱导这些细胞中p53相互作用的含有WW结构域的氧化还原酶（WOX1，也称为WWOX或FOR）的表达。WOX1增强肿瘤坏死因子的细胞毒功能，与p53协同介导细胞凋亡。因此，激活的JNK1可能在介导细胞凋亡中抵消WOX1。本研究表明，异位JNK1抑制wox1介导的L929成纤维细胞、单核细胞U937细胞和其他细胞类型的凋亡。此外，JNK1阻断了WOX1对细胞周期进程的预防作用。通过用大霉素或紫外光刺激细胞，JNK1被激活，WOX1在Tyr位点磷酸化（33）。通过共免疫沉淀确定，激活的JNK1与磷酸化的WOX1发生物理相互作用。WOX1中Tyr（33）到Arg（33）的改变使其与JNK1的结合相互作用及其介导细胞死亡的活性失效，表明激活WOX1需要Tyr（33）磷酸化。研究发现，显性阴性WOX1可阻断p53介导的细胞凋亡和异霉素介导的WOX1磷酸化，但不能抑制JNK1的激活。该突变蛋白结合p53，但不能与JNK1相互作用，在酵母双杂交分析中确定。综上所述，JNK1和WOX1的磷酸化对于它们的物理相互作用和功能拮抗是必要的。
Transient activation of c-Jun N-terminal kinase (JNK) promotes cell survival, whereas persistent JNK activation induces apoptosis. Bovine testicular hyaluronidase PH-20 activates JNK1 and protects L929 fibroblasts from staurosporine-mediated cell death. PH-20 also induces the expression of a p53-interacting WW domain-containing oxidoreductase (WOX1, also known as WWOX or FOR) in these cells. WOX1 enhances the cytotoxic function of tumor necrosis factor and mediates apoptosis synergistically with p53. Thus, the activated JNK1 is likely to counteract WOX1 in mediating apoptosis. Here it is demonstrated that ectopic JNK1 inhibited WOX1-mediated apoptosis of L929 fibroblasts, monocytic U937 cells, and other cell types. Also, JNK1 blocked WOX1 prevention of cell cycle progression. By stimulating cells with anisomycin or UV light, JNK1 became activated, and WOX1 was phosphorylated at Tyr(33). The activated JNK1 physically interacted with the phosphorylated WOX1, as determined by co-immunoprecipitation. Alteration of Tyr(33) to Arg(33) in WOX1 abrogated its binding interaction with JNK1 and its activity in mediating cell death, indicating that Tyr(33) phosphorylation is needed to activate WOX1. A dominant negative WOX1 was developed and shown to block p53-mediated apoptosis and anisomycin-mediated WOX1 phosphorylation but could not inhibit JNK1 activation. This mutant protein bound p53 but could not interact with JNK1, as determined in yeast two-hybrid analysis. Taken together, phosphorylation of JNK1 and WOX1 is necessary for their physical interaction and functional antagonism.