U19/Eaf2 binds to and stabilizes von hippel-lindau protein.
U19/Eaf2 binds to and stabilizes von hippel-lindau protein.
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DOI：
10.1158/0008-5472.can-08-2595
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发表时间：
2009-03-15
期刊：
影响因子：
11.2
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
Xiao W;Ai J;Habermacher G;Volpert O;Yang X;Zhang AY;Hahn J;Cai X;Wang Z
研究已经明确了肿瘤抑制因子pVHL在血管系统和正常精子发生中的关键调控作用。在这里，我们报道了敲除新发现的肿瘤抑制因子U19/Eaf2也会导致血管系统异常和曲霉发生，这表明U19/Eaf2与pVHL之间存在潜在的联系。共免疫沉淀和体外结合实验显示U19/Eaf2与pVHL之间存在关联，而缺失突变表明U19/Eaf2的NH2端和pVHL的α和β结构域都需要这种结合。U19/Eaf2对pVHL具有稳定作用，蛋白稳定性和脉冲追踪研究表明。U19/Eaf2敲除小鼠的睾丸和小鼠胚胎成纤维细胞（MEF）表达pVHL水平降低，表明pVHL在体内的完全表达确实需要U19/Eaf2。正如预期的那样，U19/Eaf2敲除MEF细胞表现出缺氧诱导因子1α (HIF1α)的水平和活性增加，HIF1α是一种通常通过pvhl介导的降解途径调节的蛋白质。此外，在Matrigel plug实验中，U19/Eaf2基因敲除小鼠的血管生成显著增加。上述观察结果表明，U19/Eaf2可能通过直接结合和稳定pVHL来调节HIF1α和血管生成。
Studies have firmly established a key regulatory role for the tumor suppressor pVHL in the regulation of the vascular system and normal spermatogenesis. Here, we report that knockout of the newly identified tumor suppressor U19/Eaf2 also caused vascular system abnormalities and aspermatogenesis, suggesting a potential link between U19/Eaf2 and pVHL. Coimmunoprecipitation and in vitro binding assays showed an association between U19/Eaf2 and pVHL, whereas deletion mutagenesis revealed the requirement of the NH2 terminus of U19/Eaf2 and both the α and β domains of pVHL for this binding. U19/Eaf2 stabilizes pVHL, as shown by protein stability and pulse-chase studies. Testes and mouse embryonic fibroblasts (MEF) derived from U19/Eaf2 knockout mice expressed reduced levels of pVHL, indicating that full in vivo expression of pVHL indeed requires U19/Eaf2. As expected, U19/Eaf2 knockout MEF cells exhibited an increased level and activity of hypoxia-inducible factor 1α (HIF1α), a protein typically regulated via a pVHL-mediated degradation pathway. Furthermore, angiogenesis in a Matrigel plug assay was significantly increased in U19/Eaf2 knockout mice. The above observations argue that U19/Eaf2 can modulate HIF1α and angiogenesis, possibly via direct binding and stabilization of pVHL.