Complex interaction between dengue virus replication and expression of miRNA-133a

Abstract

BackgroundDengue virus (DENV) is the most common vector-borne viral infection worldwide with approximately 390 million cases and 25,000 reported deaths each year. MicroRNAs (miRNAs) are small non-coding RNA molecules responsible for the regulation of gene expression by repressing mRNA translation or inducing mRNA degradation. Although miRNAs possess antiviral activity against many mammalian-infecting viruses, their involvement in DENV replication is poorly understood.MethodsHere, we explored the relationship between DENV and cellular microRNAs using bioinformatics tools. We overexpressed miRNA-133a in Vero cells to test its role in DENV replication and analyzed its expression using RT-qPCR. Furthermore, the expression of polypyrimidine tract binding protein (PTB), a protein involved in DENV replication, was analyzed by western blot. In addition, we profiled miRNA-133a expression in Vero cells challenged with DENV-2, using Taqman miRNA.ResultsBioinformatic analysis revealed that the 3' untranslated region (3'UTR) of the DENV genome of all four DENV serotypes is targeted by several cellular miRNAs, including miRNA-133a. We found that overexpression of synthetic miRNA-133a suppressed DENV replication. Additionally, we observed that PTB transcription , a miRNA-133a target, is down-regulated during DENV infection. Based in our results we propose that 3'UTR of DENV down-regulates endogenous expression of miRNA-133a in Vero cells during the first hours of infection.ConclusionsmiRNA-133a regulates DENV replication possibly through the modulation of a host factor such as PTB. Further investigations are needed to verify whether miRNA-133a has an anti-DENV effect in vivo.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-016-1364-y) contains supplementary material, which is available to authorized users.

Zika virus alters the microRNA expression profile and elicits an RNAi response in Aedes aegypti mosquitoes

“…Previously, targeting of genomes of RNA viruses by host miRNAs have been reported in mammalian cells [ 41 ]. In particular, a number of human miRNAs (hsa-miR-133a, hsa-miR-548g-3p, hsa-miR-223) with potential binding sites in the 5’ and 3’UTRs of different DENV serotypes have been shown to negatively affect replication of the viruses when overexpressed in mammalian cells [ 42 , 43 ]. In mosquitoes, a midgut-specific alb-miR-281 from Ae .…”

Letter to the Editor "Drosha, DGCR8, and Dicer mRNAs are downregulated in human cells infected with dengue virus 4" - Genet. Mol. Res. 15 (2): gmr.15027891 - Drosha, Dicer, and TRBP mRNA are downregulated in Vero cells with the 3'UTR of Dengue virus

“…We recently described a complex interaction between miR-133a and DENV replication (Castillo et al, 2016), in which we demonstrated that overexpression of miR-133a impairs DENV-2 replication in Vero cells. We also found that the 3'-untranslated regions (3'-UTRs) of DENV-1, -2, and -4 downregulated the expression of miR-133a.…”

“…In the context of this study, our data was similar to that reported by Casseb et al (2016), indicating that transfection of the 3'-UTRs of DENV-1, -2, and -4 into Vero cells induced downregulation of the expressions of Drosha, Dicer, and TAR RNA-binding protein (TRBP), all of which are key components of the miRNA biogenesis machinery. The detailed methods for obtaining the 3'-UTRs of DENV-1, -2, and -4 fused with green fluorescent protein have been described previously (Castillo et al, 2016). Vero cells were transfected with the different 3'-UTRs and total RNA was purified.…”

Avances en la investigación del virus dengue en Colombia: papel de los microARNs celulares en la respuesta anti-dengue virus

“…El primero de esos miR es muy interesante ya que está implicado en el control de la expresión de la proteína conocida como PTB (del inglés polypyrimidine tract-binding protein), la cual se ha reportado es esencial para la replicación del DENV 24 . Nuestros estudios permitieron demostrar efectivamente que la sobre-expresión del miR-133a inhibía la replicación del DENV-2 25 . Además pudimos demostrar que la infección por el DENV-2 incrementa la expresión de PTB durante las primeras horas de la infección, pero al mismo tiempo, la presencia del miR-133a contrarrestaba dicho efecto durante las primeras 12 h. Además se pudo observar que el DENV-2 inhibía la expresión endógena del miR-133a durante las primeras 8 h de la infección.…”