MiR-103a-3p Promotes Zika Virus Replication by Targeting OTU Deubiquitinase 4 to Activate p38 Mitogen-Activated Protein Kinase Signaling Pathway

Abstract

BackgroundMicroRNAs (miRNAs) play critical roles in regulating virus infection and replication. However, the mechanism by which miRNA regulates Zika virus (ZIKV) replication remains elusive. We aim to explore how the differentially expressed miR-103a-3p regulates ZIKV replication and to clarify the underlying molecular mechanism.MethodsSmall RNA sequencing (RNA-Seq) was performed to identify differentially expressed miRNAs in A549 cells with or without ZIKV infection and some of the dysregulated miRNAs were validated by quantitative real time PCR (qRT-PCR). The effect of miR-103a-3p on ZIKV replication was examined by transfecting miR-103a-3p mimic or negative control (NC) into A549 cells with or without p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 and expression levels of ZIKV NS5 mRNA and NS1 protein were detected by qRT-PCR and Western blot, respectively. The potential target genes for miR-103a-3p were predicted by four algorithms and further validated by mutation analysis through luciferase reporter assay. The predicated target gene OTU deubiquitinase (DUB) 4 (OTUD4) was over-expressed by plasmid transfection or silenced by siRNA transfection into cells prior to ZIKV infection. Activation status of p38 MAPK signaling pathway was revealed by looking at the phosphorylation levels of p38 (p-p38) and HSP27 (p-HSP27) by Western blot.ResultsThirty-five differentially expressed miRNAs in ZIKV-infected A549 cells were identified by RNA-Seq analysis. Five upregulated and five downregulated miRNAs were further validated by qRT-PCR. One of the validated upregulated miRNAs, miR-103a-3p significantly stimulated ZIKV replication both at mRNA (NS5) and protein (NS1) levels. We found p38 MAPK signaling was activated following ZIKV infection, as demonstrated by the increased expression of the phosphorylation of p38 MAPK and HSP27. Blocking p38 MAPK signaling pathway using SB203580 inhibited ZIKV replication and attenuated the stimulating effect of miR-103a-3p on ZIKV replication. We further identified OTUD4 as a direct target gene of miR-103a-3p. MiR-103a-3p over-expression or OTUD4 silencing activated p38 MAPK signaling and enhanced ZIKV replication. In contrast, OTUD4 over-expression inhibited p38 MAPK activation and decreased ZIKV replication. In addition, OTUD4 over-expression attenuated the stimulating effect of miR-103a-3p on ZIKV replication and activation of p38 MAPK signaling.ConclusionZika virus infection induced the expression of miR-103a-3p, which subsequently activated p38 MAPK signaling pathway by targeting OTUD4 to facilitate ZIKV replication.

Protective Effects of Caffeine on Chikungunya and Zika Virus Infections: An in Vitro and in Silico Study

“…Although little is known about the interactions between ZIKV infection and host metabolism, previous studies reported that ZIKV also takes advantage of the p38 AMPK signaling. Indeed, ZIKV infection leads to the activation of p38 MAPK signaling, as was demonstrated by increased phosphorylation of p38 MAPK and blocking of MAPK signaling during ZIKV replication [48] . In addition, ZIKV also induces the activation of JNK and p38 MAPK in trophoblasts after 16 to 24 h of infection [49] .…”

“…Indeed, ZIKV infection leads to the activation of p38 MAPK signaling, as was demonstrated by increased phosphorylation of p38 MAPK and blocking of MAPK signaling during ZIKV replication. [48] In addition, ZIKV also induces the activation of JNK and p38 MAPK in trophoblasts after 16 to 24 h of infection. [49] Thus, caffeine could act antagonistically and target MAPK and JNK signaling, which would explain its antiviral effect against CHIKV and ZIKV as reported here.…”

Integrated analysis to study the interplay between post-translational modifications (PTM) in hepatitis C virus proteins and hepatocellular carcinoma (HCC) development

“…Moreover, p38 MAPK has also been previously reported to play a critical pro-viral role in the replication of several other viruses including influenza A virus 31 , 32 , HIV 33 , Human Cytomegalovirus 34 , Varicella-Zoster 35 , Chikungunya Virus 36 and Junin virus 37 . On the other hand, inhibition of p38 signaling has been reported to impair replication of flaviviruses, respiratory viruses, enterovirus, rotaviruses, Chikungunya virus, SARS-CoV-2, junin virus as well as zika virus 37 , 38 . All these studies support the conclusion that these kinases are tightly involved in the positive regulation of HCV life cycle.…”

OTUD4 regulates metastasis and chemoresistance in melanoma by stabilizing Snail1

“…For instance, OTUD4 is downregulated by the ZIKV infection induced miR‐103a‐3p to stimulate replication (Liuyu et al, 2019). In addition, viral infection was reported to promote its transcriptional activation by IRF3/7‐binding on the promoter of Otud4 (Ye et al, 2022). High level transcription of OTUD4 was significantly observed in tsc22d3‐overexpressing embryos in Bmp‐dependent dorsoventral patterning (Tse et al, 2013).…”