Hyaluronidase induction of a WW domain-containing oxidoreductase that enhances tumor necrosis factor cytotoxicity
Hyaluronidase induction of a WW domain-containing oxidoreductase that enhances tumor necrosis factor cytotoxicity
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DOI：
10.1074/jbc.m007140200
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发表时间：
2001-02-02
影响因子：
4.8
通讯作者：
Zevotek, N
中科院分区：
文献类型：
--
作者：
Chang, NS;Pratt, N;Zevotek, N
为了确定透明质酸酶如何增加某些癌细胞对肿瘤坏死因子（TNF）细胞毒性的敏感性，我们在这里报道了透明质酸酶诱导的小鼠含WW结构域的氧化还原酶（WOX 1）的分离和表征，WOX 1由两个N-末端WW结构域、一个核定位序列和一个C-末端醇脱氢酶（ADH）结构域组成。WOX 1主要定位于线粒体中，线粒体靶向序列定位于ADH结构域内。TNF、staurosporine和甘草酸苷诱导线粒体通透性转换导致WOX 1从线粒体释放并随后发生核转位，TNF介导的WOX 1核转位发生在核因子-κ B核转位之后不久，但两者都是独立的事件。如使用稳定的细胞转染子所确定的，W 0X 1通过其WW和ADH结构域增强L929细胞中的TNF细胞毒性。与此观察平行，W 0X 1还在瞬时表达实验中增强TRADD（TNF受体相关死亡结构域蛋白）介导的细胞死亡。反义表达WOX 1可提高L929细胞对TNF的抵抗力。WOX 1增强TNF细胞毒性部分是由于其显著下调凋亡抑制剂Bcl-2和Bcl-x（L）（>85%），但通过ADH结构域上调促凋亡p53（类似于200%）。当过表达时，ADH结构域介导的细胞凋亡，可能是由于这些蛋白质的表达的调制。WW结构域不能调节这些蛋白的表达，但使COS-7细胞对TNF杀伤敏感，并独立于半胱天冬酶介导各种癌细胞的凋亡。瞬时共转染细胞与p53和WOX 1以协同方式诱导凋亡。WOX 1与p53共定位于胞质溶胶中，并通过其WW结构域结合到p53的富含脯氨酸的区域。通过反义mRNA阻断WOX 1的表达可阻断p53的凋亡。因此，WOX 1是一种线粒体致凋亡蛋白，是p53在细胞死亡中的重要伙伴。
To determine how hyaluronidase increases certain cancer cell sensitivity to tumor necrosis factor (TNF) cytotoxicity, we report here the isolation and characterization of a hyaluronidase-induced murine WW domain-containing oxidoreductase (WOX1), WOX1 is composed of two N-terminal WW domains, a nuclear localization sequence, and a C-terminal alcohol dehydrogenase (ADH) domain. WOX1 is mainly located in the mitochondria, and the mitochondrial targeting sequence was mapped within the ADH domain. Induction of mitochondrial permeability transition by TNF, staurosporine, and atractyloside resulted in WOX1 release from mitochondria and subsequent nuclear translocation, TNF-mediated WOX1 nuclear translocation occurred shortly after that of nuclear factor-kappaB nuclear translocation, whereas both were independent events. WOX1 enhanced TNF cytotoxicity in L929 cells via its WW and ADH domains as determined using stable cell transfectants, In parallel with this observation, WOX1 also enhanced TRADD (TNF receptor-associated death domain protein)-mediated cell death in transient expression experiments. Antisense expression of WOX1 raised TNF resistance in L929 cells. Enhancement of TNF cytotoxicity by WOX1 is due, in part, to its significant downregulation of the apoptosis inhibitors Bcl-2 and Bcl-x(L) (>85%), but up-regulation of pro-apoptotic p53 (similar to 200%) by the ADH domain. When overexpressed, the ADH domain mediated apoptosis, probably due to modulation of expression of these proteins. The WW domains failed to modulate the expression of these proteins, but sensitized COS-7 cells to TNF killing and mediated apoptosis in various cancer cells independently of caspases, Transient cotransfection of cells with both p53 and WOX1 induced apoptosis in a synergistic manner. WOX1 colocalizes with p53 in the cytosol and binds to the proline-rich region of p53 via its WW domains. Blocking of WOX1 expression by antisense mRNA abolished p53 apoptosis, Thus, WOX1 is a mitochondrial apoptogenic protein and an essential partner of p53 in cell death.