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

Abstract

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. IMPORTANCE This 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.

Stress proteins: the biological functions in virus infection, present and challenges for target-based antiviral drug development

“…RNA virus: YFV 350 Immunity modulation RNA virus: HFDV 354 Cellular transformation DNA virus: HBV, [363][364][365][366] Small chaperone Hsp27 Viral replication RNA virus: EV-A71, 384 Swine fever virus (CSFV), 388 Porcine epidemic diarrhoea virus (PEDV) 389 DNA virus: Adenovirus, 390 HSV-1, 397 Virus entry and uncoating RNA virus: Newcastle disease Virus, 402 DEGV [402][403][404] DNA virus: SV40 413 RT virus: HIV 407,410 Virus translation RNA virus: EV-A71 415 Oxidative stress and ER stress RNA virus: Influenza, 419 HCV, 421 EMCV, 422 435 Coronavirus, 436 Poliovirus, 438 HCV 437 DNA virus: HCMV 488 RNA transcription DNA virus: HPV18, 425,489 HBV, 490 HSV 492 RT virus: HIV 514 RNA polyadenylation RT virus: Rous sarcoma virus 485,486 (HDAC6/8) in the influenza RdRp nuclear import stage. HDAC6/8 inhibitors efficiently limit the polymerase nuclear import and suppress virus replication.…”

Foot-and-Mouth Disease Virus Capsid Protein VP1 Interacts with Host Ribosomal Protein SA To Maintain Activation of the MAPK Signal Pathway and Promote Virus Replication

“…To better understand the potential pathogenic consequences of VP1-host protein interactions, we performed a yeast two-hybrid (YTH) assay as previously described and reported that VP1 interacted with host DnaJ heat shock protein family member A3 (DNAJA3) and showed that DNAJA3 played an essential role for suppression of FMDV replication (7). In the present study, we report that FMDV VP1 interacted with host ribosomal protein SA (RPSA), a component of the ribosome (8,9).…”

“…FMDV structural VP1 protein interacted with porcine host protein RPSA. Cellular RPSA was identified as a potential target of FMDV VP1 by YTH assay in our previous study (7). To confirm the VP1-RPSA interaction, we performed the coimmunoprecipitation assay.…”

“…Moreover, VP1 inhibits IRF3 phosphorylation to impair type I interferon production and promote virus replication. The host DNAJA3 protein also interacts with the C-terminal 188 -211 region of VP1 to induce the lysosomal degradation of VP1 and attenuate VP1-mediated suppressive effect on interferon production (7). The 135-144 region of FMDV VP1 contains both Band T-cell epitopes and has been used as an immunogen in hosts (33).…”

Advances in Foot-and-Mouth Disease Virus Proteins Regulating Host Innate Immunity

“…Recently, the interaction of the host cell protein DnaJ heat shock protein family (Hsp40) member A3 (DNAJA3) with VP1 was identified through a yeast two-hybrid system. Studies has shown that DNAJA3 can significantly reduce the inhibitory effect of VP1 on SeV-induced IRF3 phosphorylation, dimerization, and nuclear localization through the lysosomal pathway, thus reducing the antagonism on the IFN-β signal pathway and inhibiting FMDV replication ( Zhang et al, 2019 ). In addition, VP1 can also act on ribosomal protein SA (RPSA) to weaken its inhibitory effect on the mitogen-activated protein kinase (MAPK) pathway, which is conducive to the FMDV replication ( Zhu et al, 2020 ).…”