MiR-23a Contributes HSV-1 Replication And Its Mechanism

Objective:MicroRNAs(miRNAs)are a class of small non-coding RNAs including 19-22 nucleotides,depending on the complementarity base-pairing,which act as specific post-transcriptional inhibitors of mRNA function in cellular self-regulation.And these small non-coding RNAs can also contribute to the repertoire of host-pathogen complex interactions during viral infection.Host-cellular miRNAs reported that can modulate the expression of various viral genes against viral infection;several viral miRNAs have been shown to inhibit the expression of cellular factors that play a role in cellular innate antiviral immune responses;even viruses are capable of using cellular miRNAs to promote their replication.In this study,we focus on the effects of miR-23a on the replication of HSV-1 in HeLa cells,as well as the identification of the direct target genes,in order to illuminate the possible mechanism of miR-23a in HSV-1 replication.Methods:After miR-23a was ectopic expressed or depleted by transfected with pcDNA3/miR-23a or pRNAT-U6.2/miR-23a,we used standard plaque assay,neutral red staining and real-time PCR to explore the role of miR-23a in virus replication.According to the result that miR-23a could facilitates HSV-1 replication in HeLa cells,next we identified the candidate target genes for miR-23a combined with the bioinformation and previous study.And then we focused on IRF1 gene(Interferon Regulatory Factor 1).The result of fluorescent report assay suggested that in HeLa cells,miR-23a could directly bind to IRF1 3' UTR and negatively regulated IRF1 expression.Furthermore,in order to confirm the regulating role of miR-23a in target gene IRF1 expression,the mRNA levels and protein levels of IRF1 were detected with real-time RT-PCR and western blot,which was consistent with the expected.Subsequently,contrary to the function of miR-23a,we determined that IRF1 suppressed HSV-1 replication in HeLa cells.With the help of co-transfected with pcDNA3/miR-23a and IRF1,we found that present of IRF1 could weaken the strong pro-viral effect induced by miR-23 a.Results:1.MiR-23a facilitates HSV-1 replication in HeLa cells.2.MiR-23a targets IRF1 directly and negatively regulates IRF1 expression in HeLa cells.3.IRF1 gene confers an antiviral state in HeLa cells infected with HSV-1.4.Present of IRF1 can weaken the strong pro-viral effect induced by miR-23a.5.HSV-1 infection of HeLa cells induces miRNA-23a and restrains IRF16.IRF-1 inhibits virus replication through induction of viperin expression.Conclusions:Over-expression of miR-23a led to a pro-effect on HSV-1 replication,such as promoting viral DNA synthesis and increasing the concentration of infectious viruses in the supernatant.IRF1 serves as a transcription activator,which plays an important role in innate immune system.We found that miR-23a targeted IRF1 directly and negatively regulated IRF1 expression in HeLa cells,and the present of IRF1 could weaken the strong pro-viral effect induced by miR-23a.Furthermore,IRF-1 inducted viperin expression to inhibit virus replication through.Depending on this study,we inferred that IRF1 mediated the miR-23a exerting a strong pro-viral response,which enable us to understand the host-virus interaction better.