Human miR-3145 inhibits influenza A viruses replication by targeting and silencing viral PB1 gene

Abstract

MicroRNAs (miRNAs) play an important role in the regulation of gene expression and are involved in many cellular processes including inhibition of viral replication in infected cells. In this study, three subtypes of influenza A viruses (pH1N1, H5N1 and H3N2) were analyzed to identify candidate human miRNAs targeting and silencing viral genes expression. Candidate human miRNAs were predicted by miRBase and RNAhybrid based on minimum free energy (MFE) and hybridization patterns between human miRNAs and viral target genes. In silico analysis presented 76 miRNAs targeting influenza A viruses, including 70 miRNAs that targeted specific subtypes (21 for pH1N1, 27 for H5N1 and 22 for H3N2) and 6 miRNAs (miR-216b, miR-3145, miR-3682, miR-4513, miR-4753 and miR-5693) that targeted multiple subtypes of influenza A viruses. Interestingly, miR-3145 is the only candidate miRNA targeting all three subtypes of influenza A viruses. The miR-3145 targets to PB1 encoding polymerase basic protein 1, which is the main component of the viral polymerase complex. The silencing effect of miR-3145 was validated by 3 0 -UTR reporter assay and inhibition of influenza viral replication in A549 cells. In 3 0 -UTR reporter assay, results revealed that miR-3145 triggered significant reduction of the luciferase activity. Moreover, expression of viral PB1 genes was also inhibited considerably (P value < 0.05) in viral infected cells expressing mimic miR-3145. In conclusion, this study demonstrated that human miR-3145 triggered silencing of viral PB1 genes and lead to inhibition of multiple subtypes of influenza viral replication. Therefore, hsa-miR-3145 might be useful for alternative treatment of influenza A viruses in the future.

MicroRNA-134 regulates poliovirus replication by IRES targeting

“…1 ), suggesting an alternative mechanism of regulation. Changes in viral replication upon miRNA modulation were modest but consequential as has been shown with several other studies 35 , 36 , 57 – 60 . In this study, we show that miR-134 interacts with nts 472–492 in the domain V of Sabin-1 5′UTR.…”

MicroRNA Regulation of RNA Virus Replication and Pathogenesis

“…miRNAs can also bind to negative-strand RNA viruses after transcription of the negative-strand RNA genome into the positive-sense RNA intermediate needed for virus replication. For instance, miR-323, miR-491, miR-485, miR-654, and miR-3145 have been shown to bind to the coding region of the influenza PB1 gene segment leading to RNA degradation in infected human and canine epithelial cell lines in vitro 12 , 33 , 34 . Similar degradation of virus RNA was found when the miRNA let-7c bound to the 3′ NTR of the influenza matrix protein in human alveolar epithelial A549 cells [35] .…”

MicroRNA hsa-miR-324-5p Suppresses H5N1 Virus Replication by Targeting the Viral PB1 and Host CUEDC2

“…In contrast, another report showed that H5N1 downregulated miRNAs, such as miR-584-5p and miR-1249, that target the PB2 gene of the virus and inhibit viral replication (6). In another study, miR-3145 inhibited influenza virus replication by targeting and silencing the PB1 gene (43). All together, these studies along with our results indicate that the host utilizes miRNAs to restrict influenza virus via targeting the most essential genes encoding subunits (PB1 and PB2) of the viral RNA polymerase and that are indispensable for viral replication, suggesting that miRNAs are pivotal in innate immune defense against influenza virus and may be involved in restriction of other RNA viruses.…”