State Key Laboratory of Virology, School of Basic Medical Sciences, Wuhan University, Wuhan, 430072, China.
Hubei Province Key Laboratory of Allergy and Immunology, Wuhan, 430071, China.
Virol Sin. 2021 Aug;36(4):692-705. doi: 10.1007/s12250-020-00343-x. Epub 2021 Feb 3.
In recent years, various serious diseases caused by Zika virus (ZIKV) have made it impossible to be ignored. Confirmed existence of ZIKV in semen and sexually transmission of ZIKV suggested that it can break the blood-testis barrier (BTB), or Sertoli cell barrier (SCB). However, little is known about the underlying mechanism. In this study, interaction between actin, an important component of the SCB, and ZIKV envelope (E) protein domain III (EDIII) was inferred from co-immunoprecipitation (Co-IP) liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Confocal microscopy confirmed the role of actin filaments (F-actin) in ZIKV infection, during which part of the stress fibers, the bundles that constituted by paralleled actin filaments, were disrupted and presented in the cell periphery. Colocalization of E and reorganized actin filaments in the cell periphery of transfected Sertoli cells suggests a participation of ZIKV E protein in ZIKV-induced F-actin rearrangement. Perturbation of F-actin by cytochalasin D (CytoD) or Jasplakinolide (Jas) enhanced the infection of ZIKV. More importantly, the transepithelial electrical resistance (TEER) of an in vitro mouse SCB (mSCB) model declined with the progression of ZIKV infection or overexpression of E protein. Co-IP and confocal microscopy analyses revealed that the interaction between F-actin and tight junction protein ZO-1 was reduced after ZIKV infection or E protein overexpression, highlighting the role of E protein in ZIKV-induced disruption of the BTB. We conclude that the interaction between ZIKV E and F-actin leads to the reorganization of F-actin network, thereby compromising BTB integrity.
近年来，寨卡病毒（ZIKV）引起的各种严重疾病已不容忽视。寨卡病毒在精液中被证实存在，且寨卡病毒可通过性传播，这表明它可以破坏血睾屏障（BTB）或支持细胞屏障（SCB）。然而，其潜在机制仍知之甚少。在这项研究中，通过共免疫沉淀（Co-IP）液相色谱-串联质谱（LC-MS/MS）分析推断出， actin（SCB 的重要组成部分）与寨卡病毒包膜（E）蛋白结构域 III（EDIII）之间的相互作用。共聚焦显微镜证实了肌动蛋白丝（F-actin）在寨卡病毒感染中的作用，在此过程中，部分应激纤维（由平行肌动蛋白丝组成的束）被破坏并出现在细胞外周。转染支持细胞中 E 和重新组织的肌动蛋白丝在细胞外周的共定位表明，寨卡病毒 E 蛋白参与了寨卡病毒诱导的 F-actin 重排。细胞松弛素 D（CytoD）或 Jasplakinolide（Jas）对 F-actin 的扰动增强了寨卡病毒的感染。更重要的是，体外小鼠 SCB（mSCB）模型的跨上皮电阻（TEER）随着寨卡病毒感染或 E 蛋白过表达的进展而下降。Co-IP 和共聚焦显微镜分析表明，寨卡病毒感染或 E 蛋白过表达后，F-actin 与紧密连接蛋白 ZO-1 的相互作用减少，突出了 E 蛋白在寨卡病毒诱导的 BTB 破坏中的作用。我们的结论是，寨卡病毒 E 与 F-actin 之间的相互作用导致 F-actin 网络的重组，从而破坏 BTB 的完整性。
