MicroRNA-33a disturbs influenza A virus replication by targeting ARCN1 and inhibiting viral ribonucleoprotein activity

Abstract

In order to explore the roles of microRNA(s) [miRNA(s)] in the influenza A virus life cycle, we compared the miRNA profiles of 293T and HeLa cell lines, as influenza A virus can replicate efficiently in 293T cells but only poorly in HeLa cells. We analysed differentially expressed miRNAs and identified five, including miR-33a, that could disturb influenza A virus replication significantly. Using TargetScan analysis, we found that ARCN1 could be a potential target of miR-33a. To confirm whether miR-33a could truly target ARCN1, we generated a luciferase reporter for the ARCN1 39 untranslated region (UTR) and performed a luciferase assay. The data indicated that miR-33a could suppress the luciferase activity of the reporter for the ARCN1 39 UTR but not a reporter in which the predicted miR-33a targeting sites on ARCN1 39 UTR were mutated. We performed immunoblotting to confirm that miR-33a could downregulate the protein level of ARCN1. Consistently, the level of ARCN1 protein in HeLa cells was significantly lower than that in 293T cells. We also demonstrated that ectopic expression of ARCN1 could partially rescue the inhibitory effect of miR-33a on virus replication. Furthermore, we demonstrated that miR-33a could impede virus replication at the stage of virus internalization, which was similar to the pattern for knockdown of ARCN1, indicating that miR-33a inhibits influenza virus infection by suppressing ARCN1 expression. In addition, we found that miR-33a could also weaken the viral ribonucleoprotein activity in an ARCN1-independent manner. In conclusion, we found that miR-33a is a novel inhibitory factor for influenza A virus replication.

The role of miR‐146a in viral infection

“…Previous evidence has demonstrated that several miRNAs, including miR‐33a, miR‐302c, and miR‐let‐7c, are involved in the replication of IAV . It has also been shown that miR‐146a is induced via a wide range of viral infections, and probably miR‐146a contributes to virus replication.…”

“…50 Previous evidence has demonstrated that several miRNAs, including miR-33a, miR-302c, and miR-let-7c, are involved in the replication of IAV. [99][100][101] It has also been shown that miR-146a is induced via a wide range of viral infections, and probably miR-146a contributes to virus replication. However, studies on the involvement of miR-146a in IAV infection are insufficient, and the related underlying molecular mechanisms of influenza infectious cycle remain unknown.…”

Human microRNA‐30 inhibits influenza virus infection by suppressing the expression of SOCS1, SOCS3, and NEDD4

“…Many studies have demonstrated the significant effect of miRNAs on modulating influenza virus infection. For examples, miR‐33a was suggest to disrupt influenza A virus (IAV) replication by targeting APCN1 (Hu et al, ); miR‐34a contributed to influenza virus‐mediated apoptosis by binding to BAX (Fan & Wang, ); and miR‐let‐7c targeted and inhibited M1 expression of the H1N1 influenza virus (Ma et al, ). Although several studies linked miRNAs to type I IFN production mediated by RIG‐I in influenza virus infection (miR‐144, miR‐485, and miR‐136) (Ingle et al, ; Rosenberger et al, ; Zhao et al, ), the underlying regulation of the type I IFN‐mediated antiviral response by miRNAs during influenza virus infection remains unclear.…”

Cellular microRNA miR-c89 inhibits replication of porcine reproductive and respiratory syndrome virus by targeting the host factor porcine retinoid X receptor β

“…It has been reported that a liver-specific microRNA, miR-122, can promote the replication of hepatitis C virus (HCV) [13,14], while miR-199a inhibits viral replication by binding to HCV genomic RNA [15]. Similarly, miR-33a can inhibit viral replication in influenza A by suppressing ARCN1 expression and weakening viral ribonucleoprotein activity [16], and miR-32 plays a crucial role in a the antiviral response to primate foamy virus type 1 (PFV-1) by targeting ORF2 of the viral genome [17]. Additionally, the EBNA1 protein of Epstein-Barr virus (EBV) promotes EBV latency by inducing the expression of let-7 miRNAs [18] and, of particular note, miR-181 inhibits PRRSV replication by targeting both viral genomic RNA and cellular receptor CD163 [19,20].…”