CK2 phosphorylation of Schistosoma mansoni HMGB1 protein regulates its cellular traffic and secretion but not its DNA transactions.
CK2 phosphorylation of Schistosoma mansoni HMGB1 protein regulates its cellular traffic and secretion but not its DNA transactions.
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曼氏菌HMGB1蛋白的CK2磷酸化调节其细胞流量和分泌，但不能调节其DNA交易。
DOI：
10.1371/journal.pone.0023572
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Fantappié MR
中科院分区：
文献类型：
--
作者：
de Abreu da Silva IC;Carneiro VC;Maciel Rde M;da Costa RF;Furtado DR;de Oliveira FM;da Silva-Neto MA;Rumjanek FD;Fantappié MR
曼氏血吸虫寄生虫寄生在肠系膜静脉中，受精的雌性蠕虫每天在那里产卵数百个。一些鸡蛋抗原被困在肝脏中，并引发强烈的肉芽肿反应。高迁移率族蛋白1(HMGB1)是一种核因子，也可以分泌并发挥细胞因子的作用。血吸虫HMGB1(SmHMGB1)由虫卵分泌，刺激参与血吸虫病病理过程的关键细胞因子的产生。因此，了解SmHMGB1的释放机制变得非常必要。在这里，我们解决了核SmHMGB1如何到达细胞外空间的问题。我们在体外和体内证明，CK2磷酸化参与了SmHMGB1的核质穿梭。通过定点突变，我们定位了被CK2磷酸化的SmHMGB1的两个丝氨酸残基。DNA弯曲和超螺旋实验表明，SmHMGB1的CK2磷酸化对该蛋白的DNA结合活性没有影响。电子显微镜、细胞转染法和荧光显微镜显示SmHMGB1存在于成体血吸虫和哺乳动物细胞的胞核和胞浆中。此外，我们还发现，无论是用磷酸酶还是用CK2抑制剂处理细胞，都能分别增强或阻断SmHMGB1的细胞流量。重要的是，我们通过共聚焦显微镜和生化方法证明了SmHMGB1是由感染动物的曼氏血吸虫虫卵显著分泌的，并且定位于卵泡周围血吸虫肉芽肿的SmHMGB1被磷酸化。我们发现SmHMGB1的分泌受磷酸化的调节。此外，我们的结果提示，虫卵分泌的SmHMGB1可能代表了一种新的虫卵抗原。因此，寻找特异性靶向SmHMGB1磷酸化的药物可能会阻断SmHMGB1的分泌并干扰血吸虫病的发病机制。
The helminth Schistosoma mansoni parasite resides in mesenteric veins where fecundated female worms lay hundred of eggs daily. Some of the egg antigens are trapped in the liver and induce a vigorous granulomatous response. High Mobility Group Box 1 (HMGB1), a nuclear factor, can also be secreted and act as a cytokine. Schistosome HMGB1 (SmHMGB1) is secreted by the eggs and stimulate the production of key cytokines involved in the pathology of schistosomiasis. Thus, understanding the mechanism of SmHMGB1 release becomes mandatory. Here, we addressed the question of how the nuclear SmHMGB1 can reach the extracellular space. We showed in vitro and in vivo that CK2 phosphorylation was involved in the nucleocytoplasmic shuttling of SmHMGB1. By site-directed mutagenesis we mapped the two serine residues of SmHMGB1 that were phosphorylated by CK2. By DNA bending and supercoiling assays we showed that CK2 phosphorylation of SmHMGB1 had no effect in the DNA binding activities of the protein. We showed by electron microscopy, as well as by cell transfection and fluorescence microscopy that SmHMGB1 was present in the nucleus and cytoplasm of adult schistosomes and mammalian cells. In addition, we showed that treatments of the cells with either a phosphatase or a CK2 inhibitor were able to enhance or block, respectively, the cellular traffic of SmHMGB1. Importantly, we showed by confocal microscopy and biochemically that SmHMGB1 is significantly secreted by S. mansoni eggs of infected animals and that SmHMGB1 that were localized in the periovular schistosomotic granuloma were phosphorylated. We showed that secretion of SmHMGB1 is regulated by phosphorylation. Moreover, our results suggest that egg-secreted SmHMGB1 may represent a new egg antigen. Therefore, the identification of drugs that specifically target phosphorylation of SmHMGB1 might block its secretion and interfere with the pathogenesis of schistosomiasis.
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影响因子：
30.8
作者：
Calogero, S;Grassi, F;Bianchi, ME
通讯作者：
Bianchi, ME
影响因子：
4.4
作者：
Duraes, Fernanda V.;Carvalho, Natalia B.;Fonseca, Cristina T.
通讯作者：
Fonseca, Cristina T.
影响因子：
4.4
作者：
Henri, S;Chevillard, C;Dessein, AJ
通讯作者：
Dessein, AJ
影响因子：
4
作者：
Davies, SJ;Lim, KC;McKerrow, JH
通讯作者：
McKerrow, JH
DOI：
10.1016/j.bbrc.2009.10.129
发表时间：
2009-12-25
影响因子：
3.1
作者：
Carneiro, Vitor Coutinho;Maciel, Renata de Moraes;Fantappie, Marcelo Rosado
通讯作者：
Fantappie, Marcelo Rosado