Deleted in azoospermia-associated protein 2 regulates innate immunity by stimulating Hippo signaling in crab
Deleted in azoospermia-associated protein 2 regulates innate immunity by stimulating Hippo signaling in crab
复制标题
无精子症相关蛋白 2 中的缺失通过刺激螃蟹中的 Hippo 信号来调节先天免疫
DOI：
10.1074/jbc.ra119.009559
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发表时间：
2019-10-04
影响因子：
4.8
通讯作者：
Li，Weiwei
中科院分区：
文献类型：
--
作者：
Yang，Lei;Li，Xuejie;Li，Weiwei
河马信号通路在正常的动物生理和发病机制中都起着关键作用。由于针对这一途径的药物干预具有不同的临床意义，因此更好地了解其在各种条件和生物体中的调节是至关重要的。在此，我们在中华绒螯蟹(Eriocheir Sinensis)中发现了缺失的无精子相关蛋白2(DAZAP2)，命名为EsDAZAP2，它是一种河马调节蛋白，与各种昆虫、鱼类和哺乳动物中的蛋白高度相似。我们发现，细菌感染显著诱导EsDAZAP2的表达，而EsDAZAP2基因敲除在体外既抑制了抗菌肽(AMP)的表达，又导致体内活菌数和死亡率的增加，表明EsDAZAP2在先天性免疫中起着关键作用。利用酵母双杂交筛选和免疫共沉淀实验，我们发现EsDAZAP2调控的是Toll途径，而不是免疫缺陷和Janus激酶(JAK)/信号转导和转录激活因子(STAT)途径。我们的发现还表明EsDAZAP2与河马蛋白萨尔瓦多(Sav)结合。此外，通过研究调节中华绒毛虫AMP表达的转录因子DRADAL的调控，我们提供了实验证据，表明EsDAZAP2在天然免疫中促进了河马途径的激活，EsDAZAP2和Hippo与不同的SAV结构域结合。据我们所知，这是DAZAP2调节的河马信号通路在动物先天免疫中作用的第一个报告。
The Hippo-signaling pathway plays a critical role in both normal animal physiology and pathogenesis. Because pharmacological interventions targeting this pathway have diverse clinical implications, a better understanding of its regulation in various conditions and organisms is crucial. Here, we identified deleted in azoospermia-associated protein 2 (DAZAP2) in the Chinese mitten crab (Eriocheir sinensis), designated EsDAZAP2, as a Hippo-regulatory protein highly similar to proteins in various species of insects, fish, and mammals. We found that a bacterial infection significantly induces EsDAZAP2 expression, and an EsDAZAP2 knockdown both suppresses antimicrobial peptide (AMP) expression in vitro and results in increased viable bacterial counts and mortality in vivo, suggesting that EsDAZAP2 plays a critical role in innate immunity. Using yeast two-hybrid screening and co-immunoprecipitation assays, we found that EsDAZAP2 regulates the Toll pathway rather than the immune deficiency and Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathways. Our findings also demonstrate that EsDAZAP2 binds to the Hippo protein, Salvador (Sav). Moreover, by examining the regulation of Dorsal, a transcription factor that regulates AMP expression in E. sinensis, we provide experimental evidence indicating that EsDAZAP2 promotes Hippo pathway activation in innate immunity, with EsDAZAP2 and Hippo binding to different Sav domains. To the best of our knowledge, this is the first report of a DAZAP2-regulated Hippo-signaling pathway operating in animal innate immunity.