Role of Epidermal Growth Factor Receptor Signaling in the Interaction of Neisseria meningitidis with Endothelial Cells
Role of Epidermal Growth Factor Receptor Signaling in the Interaction of Neisseria meningitidis with Endothelial Cells
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DOI：
10.1128/iai.01346-13
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发表时间：
2014-03-01
影响因子：
3.1
通讯作者：
Schubert-Unkmeir, Alexandra
中科院分区：
文献类型：
--
作者：
Slanina, Heiko;Muendlein, Sabrina;Schubert-Unkmeir, Alexandra
脑膜炎奈瑟菌是脑膜炎和败血症的病原体，附着并侵入各种细胞类型。这两个步骤都诱导和/或需要宿主细胞蛋白的酪氨酸磷酸化。在这里，我们使用磷酸化阵列平台来鉴定N.脑膜炎感染的脑内皮细胞来破译细菌摄取所必需的RTK依赖性信号通路。我们检测到几种激活的RTK，包括ErbB家族受体表皮生长因子受体（EGFR），ErbB 2和ErbB 4。我们发现，药物抑制和ErbB受体酪氨酸磷酸化和表达的基因消融导致中国卵巢仓鼠（CHO-K1）细胞中EGFR、ErbB 2或ErbB 4的细菌摄取和异源表达减少，而这些细胞不表达EGFR和ErbB 4;这种减少导致脑膜炎球菌侵袭显著增加。EGFR和ErbB 4的激活通过共同配体HB-EGF（肝素结合EGF样配体）的反式激活介导，其在感染的细胞培养上清液中显著升高。我们进一步确定N.脑膜炎诱导EGFR在Tyr 845处的磷酸化，而不依赖于配体结合，这需要c-Src活化并参与介导N的摄取。将脑膜炎病毒转化为真核细胞。EGFR Y845 F的表达抑制了摄取增加，EGFR Y845 F在激酶结构域中具有点突变。此外，ErbB 4在其自身磷酸化位点Tyr 1284处的活化和ErbB 2 Thr 686的磷酸化被观察到。总之，我们的研究结果提供了证据，EGFR，ErbB 2和ErbB 4被激活响应N。脑膜炎球菌感染，并阐明了ErbB信号在脑膜炎球菌感染生物学中的作用。
Neisseria meningitidis, the causative agent of meningitis and septicemia, attaches to and invades various cell types. Both steps induce and/or require tyrosine phosphorylation of host cell proteins. Here, we used a phospho array platform to identify active receptor tyrosine kinases (RTKs) and key signaling nodes in N. meningitidis-infected brain endothelial cells to decipher RTKdependent signaling pathways necessary for bacterial uptake. We detected several activated RTKs, including the ErbB family receptors epidermal growth factor receptor (EGFR), ErbB2, and ErbB4. We found that pharmacological inhibition and genetic ablation of ErbB receptor tyrosine phosphorylation and expression resulted in decreased bacterial uptake and heterologous expression of EGFR, ErbB2, or ErbB4 in Chinese ovary hamster (CHO-K1) cells, which do not express of EGFR and ErbB4; the decrease caused a significant increase in meningococcal invasion. Activation of EGFR and ErbB4 was mediated by transactivation via the common ligand HB-EGF (heparin-binding EGF-like ligand), which was significantly elevated in infected cell culture supernatants. We furthermore determined that N. meningitidis induced phosphorylation of EGFR at Tyr845 independent of ligand binding, which required c-Src activation and was involved in mediating uptake of N. meningitidis into eukaryotic cells. Increased uptake was repressed by expression of EGFR Y845F, which harbored a point mutation in the kinase domain. In addition, activation of ErbB4 at its autophosphorylation site, Tyr1284, and phosphorylation of ErbB2 Thr686 were observed. Altogether, our results provide evidence that EGFR, ErbB2, and ErbB4 are activated in response to N. meningitidis infection and shed new light on the role of ErbB signaling in meningococcal infection biology.