Abstract
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.
Highlights
Zika virus (ZIKV), one of the mosquito-borne viruses, belongs to the genus Flavivirus of the family Flaviviridae
To discover cellular interaction partners of ZIKV E protein, we affinity-isolated proteins from mouse Sertoli cells using a ZIKV EDIII-Fc chimeric protein and we examined these proteins by SDS-PAGE
Based on LC–MS/MS data, we suspected an interaction between ZIKV EDIII and actin, which was validated by a Co-IP assay
Summary
Zika virus (ZIKV), one of the mosquito-borne viruses, belongs to the genus Flavivirus of the family Flaviviridae. ZIKV infection causes a series of symptoms, ranging from mild illness (fever, joint pain) to severe clinical manifestations (multiple organ failure, meningitis). Fontes-Garfias et al (2017) reported that glycosylation of ZIKV E protein is critical for ZIKV virulence in A129 mice and is required for ZIKV infection of the vector mosquito, Aedes aegypti. A recent study by Shan et al (2020) revealed that an evolutionary mutation in the E protein (V473M) preceding the 2015 epidemic enhanced ZIKV virulence and fitness for transmission. Zhou et al (2019) showed that the interaction between ZIKV E protein and major facilitator superfamily domain-containing protein 2 (Mfsd2a) in human brain microvascular endothelial cells enhanced the ubiquitination and degradation of Mfsd2a, hindering docosahexaenoic acid intake mediated by Mfsd2a, and eventually resulting in growth restriction of the brain
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