Neisseria meningitidis causes cell cycle arrest of human brain microvascular endothelial cells at S phase via p21 and cyclin G2
Neisseria meningitidis causes cell cycle arrest of human brain microvascular endothelial cells at S phase via p21 and cyclin G2
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DOI：
10.1111/cmi.12482
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发表时间：
2016-01-01
影响因子：
3.4
通讯作者：
Schubert-Unkmeir, Alexandra
中科院分区：
文献类型：
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作者：
Oosthuysen, Wilhelm F.;Mueller, Tobias;Schubert-Unkmeir, Alexandra
微生物病原体已经发展出几种机制来调节和干扰宿主细胞周期进程。在这项研究中，我们分析了人类病原体脑膜炎奈瑟菌对脑内皮细胞系和原代脑内皮细胞细胞周期的影响。我们发现，脑膜炎奈瑟菌在感染后3小时和24小时出现S期细胞积聚，并与G2/M期细胞减少相平行。重要的是，菌落不透明相关蛋白(OPA)家族的外膜蛋白以及OPC蛋白被证明触发了细胞在S期的积累。基于细胞周期逆转录定量聚合酶链式反应的阵列和整合网络分析揭示了几种细胞周期调控基因mRNAs丰度的变化，包括细胞周期抑制因子p21(WAF1/CIP1)和细胞周期蛋白G2。这些变化反映在脑膜炎奈瑟氏菌感染细胞的蛋白表达水平和/或重新定位的变化上。此外，p21(WAF1/CIP1)表达的增加不依赖于P53。基因消融p21(WAF1/CIP1)和细胞周期蛋白G2可阻断脑膜炎奈瑟氏菌引起的S时相积聚。最后，通过检测生物标记物8-羟基脱氧鸟苷和组蛋白变异体H_2AX的磷酸化水平，我们提供了脑膜炎奈瑟菌在感染细胞中诱导DNA氧化损伤的证据。
Microbial pathogens have developed several mechanisms to modulate and interfere with host cell cycle progression. In this study, we analysed the effect of the human pathogen Neisseria meningitidis on cell cycle in a brain endothelial cell line as well as in primary brain endothelial cells. We found that N. meningitidis causes an accumulation of cells in the S phase early at 3 and at 24 h post-infection that was paralleled by a decrease of cells in G2/M phase. Importantly, the outer membrane proteins of the colony opacity-associated (Opa) protein family as well as the Opc protein proved to trigger the accumulation of cells in the S phase. A focused cell cycle reverse transcription quantitative polymerase chain reaction-based array and integrated network analysis revealed changes in the abundance of several cell cycle regulatory mRNAs, including the cell cycle inhibitors p21(WAF1/CIP1) and cyclin G2. These alterations were reflected in changes in protein expression levels and/or relocalization in N. meningitidis-infected cells. Moreover, an increase in p21(WAF1/CIP1) expression was found to be p53 independent. Genetic ablation of p21(WAF1/CIP1) and cyclin G2 abrogated N. meningitidis-induced S phase accumulation. Finally, by measuring the levels of the biomarker 8-hydroxydeoxyguanosine and phosphorylation of the histone variant H2AX, we provide evidence that N. meningitidis induces oxidative DNA damage in infected cells.