Cellular DNAJA3, a Novel VP1-Interacting Protein, Inhibits Foot-and-Mouth Disease Virus Replication by Inducing Lysosomal Degradation of VP1 and Attenuating Its Antagonistic Role in the Beta Interferon Signaling Pathway
Cellular DNAJA3, a Novel VP1-Interacting Protein, Inhibits Foot-and-Mouth Disease Virus Replication by Inducing Lysosomal Degradation of VP1 and Attenuating Its Antagonistic Role in the Beta Interferon Signaling Pathway
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细胞 DNAJA3 是一种新型 VP1 相互作用蛋白，通过诱导 VP1 溶酶体降解并减弱其在 β 干扰素信号通路中的拮抗作用来抑制口蹄疫病毒复制。
DOI：
10.1128/jvi.00588-19
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发表时间：
2019
影响因子：
5.4
通讯作者：
Zheng Haixue
中科院分区：
文献类型：
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作者：
Zhang Wei;Yang Fan;Zhu Zixiang;Yang Yang;Wang Zhifang;Cao Weijun;Dang Wen;Li Linlin;Mao Ruoqing;Liu Yongjie;Tian Hong;Zhang Keshan;Liu Xiangtao;Ma Junwu;Zheng Haixue
DNAJ热休克蛋白家族（Hsp 40）成员A3（DNAJA 3）在病毒感染中起重要作用。然而，DNAJA 3在口蹄疫病毒（FMDV）复制中的作用仍不清楚。在这项研究中，DNAJA 3，一个新的结合伙伴的VP 1，被确定为酵母双杂交筛选。DNAJA 3-VP 1的相互作用进一步证实了免疫共沉淀和共定位在口蹄疫病毒感染的细胞。DNAJA 3的J结构域（氨基酸1至168）和VP 1的位置208（K208）的赖氨酸显示对于DNAJA 3-VP 1相互作用是关键的。DNAJA 3的过表达显著抑制FMDV的复制，而DNAJA 3的功能丧失引起对抗FMDV复制的相反作用。机制研究表明，VP 1的K208是使用K208 A突变病毒降低DNAJA 3引起的病毒滴度的关键。DNAJA 3通过与LC 3相互作用，增强溶酶体途径的激活，诱导VP 1的溶酶体降解。同时，我们发现VP 1通过抑制IRF 3的磷酸化、二聚化和核转位来抑制β干扰素（IFN-β）信号通路。这种抑制作用在DNAJA 3敲除细胞中得到显著增强。相反，DNAJA 3的过表达显著减弱了VP 1介导的对IFN-β信号通路的抑制。与DNAJA 3-WT细胞相比，DNAJA 3-敲除细胞中Poly（I β C）诱导的IRF 3磷酸化也降低。总之，我们的研究描述了一个新的作用，DNAJA 3在宿主的抗病毒反应，诱导溶酶体降解的VP 1和减弱VP 1诱导的抑制作用的IFN-β signals.IMPORTANCEThis研究开创性地确定了DNAJA 3在口蹄疫病毒的抗病毒作用。发现DNAJA 3与FMDV VP 1相互作用并通过溶酶体途径触发其降解。此外，该研究也首次阐明了VP 1通过抑制IRF 3的磷酸化、二聚化和核转位来抑制IFN-β信号通路的机制。此外，DNAJA 3显著消除了VP 1诱导的对IFN-β信号通路的抑制作用。这些结果表明DNAJA 3通过降解VP 1和恢复IFN-β信号通路发挥抗口蹄疫病毒的作用。
DnaJ heat shock protein family (Hsp40) member A3 (DNAJA3) plays an important role in viral infections. However, the role of DNAJA3 in replication of foot-and-mouth-disease virus (FMDV) remains unknown. In this study, DNAJA3, a novel binding partner of VP1, was identified using yeast two-hybrid screening. The DNAJA3-VP1 interaction was further confirmed by coimmunoprecipitation and colocalization in FMDV-infected cells. The J domain of DNAJA3 (amino acids 1 to 168) and the lysine at position 208 (K208) of VP1 were shown to be critical for the DNAJA3-VP1 interaction. Overexpression of DNAJA3 dramatically dampened FMDV replication, whereas loss of function of DNAJA3 elicited opposing effects against FMDV replication. Mechanistical study demonstrated that K208 of VP1 was critical for reducing virus titer caused by DNAJA3 using K208A mutant virus. DNAJA3 induced lysosomal degradation of VP1 by interacting with LC3 to enhance the activation of lysosomal pathway. Meanwhile, we discovered that VP1 suppressed the beta interferon (IFN-β) signaling pathway by inhibiting the phosphorylation, dimerization, and nuclear translocation of IRF3. This inhibitory effect was considerably boosted in DNAJA3-knockout cells. In contrast, overexpression of DNAJA3 markedly attenuated VP1-mediated suppression on the IFN-β signaling pathway. Poly(I⋅C)-induced phosphorylation of IRF3 was also decreased in DNAJA3-knockout cells compared to that in the DNAJA3-WT cells. In conclusion, our study described a novel role for DNAJA3 in the host’s antiviral response by inducing the lysosomal degradation of VP1 and attenuating the VP1-induced suppressive effect on the IFN-β signaling pathway.IMPORTANCEThis study pioneeringly determined the antiviral role of DNAJA3 in FMDV. DNAJA3 was found to interact with FMDV VP1 and trigger its degradation via the lysosomal pathway. In addition, this study is also the first to clarify the mechanism by which VP1 suppressed IFN-β signaling pathway by inhibiting the phosphorylation, dimerization, and nuclear translocation of IRF3. Moreover, DNAJA3 significantly abrogated VP1-induced inhibitive effect on the IFN-β signaling pathway. These data suggested that DNAJA3 plays an important antiviral role against FMDV by both degrading VP1 and restoring of IFN-β signaling pathway.