MicroRNA-like viral small RNA from Dengue virus 2 autoregulates its replication in mosquito cells

Hussain, Mazhar and Asgari, Sassan (2014) MicroRNA-like viral small RNA from Dengue virus 2 autoregulates its replication in mosquito cells. Proceedings of the National Academy of Sciences, Early Edition 7: 1-6. doi:10.1073/pnas.1320123111

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Author Hussain, Mazhar
Asgari, Sassan
Title MicroRNA-like viral small RNA from Dengue virus 2 autoregulates its replication in mosquito cells
Journal name Proceedings of the National Academy of Sciences   Check publisher's open access policy
ISSN 0027-8424
1091-6490
Publication date 2014-02-03
Year available 2014
Sub-type Article (original research)
DOI 10.1073/pnas.1320123111
Open Access Status Not yet assessed
Volume Early Edition
Issue 7
Start page 1
End page 6
Total pages 6
Place of publication Washington, DC, United States
Publisher National Academy of Sciences
Language eng
Abstract MicroRNAs (miRNAs) are small regulatory RNAs that play significant roles in most cellular processes. In the seemingly endless arms race between hosts and pathogens, viruses also encode miRNAs that facilitate successful infection. In search of functional miRNAs or viral small RNAs (vsRNAs) encoded by Dengue virus (DENV), deep sequencing data of virus-infected Aedes aegypti mosquitoes were used. From six vsRNAs, with candidate stem-loop structures in the 5' and 3' untranslated regions of the viral genomic RNA, inhibition of DENV-vsRNA-5 led to significant increases in viral replication. Silencing of RNA interference (RNAi)/miRNA pathways' associated proteins showed that Argonaute 2 is mainly involved in DENV-vsRNA-5 biogenesis. Cloning of the precursor stem loop, immunoprecipitations, ectopic expression and detection in RNAi-deficient C6/36, and the mammalian Vero cell lines further confirmed DENV-vsRNA-5 production. Furthermore, significant impact of synthetic mimic and inhibitor of DENV-vsRNA-5 on DENV RNA levels revealed DENV-vsRNA-5's role in virus autoregulation by targeting the virus nonstructural protein 1 gene. Notably, DENV-vsRNA-5 homologous mimics from DENV serotypes 1 and 4, but not 3, inhibited DENV-2 replication. The results revealed that DENV is able to encode functional vsRNAs, and one of those, which resembles miRNAs, specifically targets a viral gene, opening an avenue for possible utilization of the small RNA to limit DENV replication.
Formatted abstract
MicroRNAs (miRNAs) are small regulatory RNAs that play significant roles in most cellular processes. In the seemingly endless arms race between hosts and pathogens, viruses also encode miRNAs that facilitate successful infection. In search of functional miRNAs or viral small RNAs (vsRNAs) encoded by Dengue virus (DENV), deep sequencing data of virus-infected Aedes aegypti mosquitoes were used. From six vsRNAs, with candidate stem-loop structures in the 5′ and 3′ untranslated regions of the viral genomic RNA, inhibition of DENV–vsRNA-5 led to significant increases in viral replication. Silencing of RNA interference (RNAi)/miRNA pathways’ associated proteins showed that Argonaute 2 is mainly involved in DENV–vsRNA-5 biogenesis. Cloning of the precursor stem loop, immunoprecipitations, ectopic expression and detection in RNAi-deficient C6/36, and the mammalian Vero cell lines further confirmed DENV–vsRNA-5 production. Furthermore, significant impact of synthetic mimic and inhibitor of DENV–vsRNA-5 on DENV RNA levels revealed DENV–vsRNA-5’s role in virus autoregulation by targeting the virus nonstructural protein 1 gene. Notably, DENV–vsRNA-5 homologous mimics from DENV serotypes 1 and 4, but not 3, inhibited DENV-2 replication. The results revealed that DENV is able to encode functional vsRNAs, and one of those, which resembles miRNAs, specifically targets a viral gene, opening an avenue for possible utilization of the small RNA to limit DENV replication.

Significance
Dengue virus has emerged as one of the most significant arboviruses, affecting millions of people around the world. Here, we show the production of a functional microRNA-like small RNA encoded by Dengue virus, which negatively regulates virus replication by targeting the viral nonstructural protein 1 sequences. The report provides a comprehensive analysis of the biogenesis and function of the viral small RNA. The results also highlight the possibility of utilizing the small RNA in inhibiting at least three serotypes of the virus. The outcomes have a significant impact on our understanding of the biology of the virus (and flaviviruses in general) and small RNA biogenesis, and open up a potential avenue for the control of the virus.
Keyword Multidisciplinary Sciences
Science & Technology - Other Topics
MULTIDISCIPLINARY SCIENCES
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID APP1027110
DE120101512
Institutional Status UQ
Additional Notes Published online before print on February 3, 2014

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
School of Biological Sciences Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 45 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 49 times in Scopus Article | Citations
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Created: Wed, 05 Feb 2014, 20:47:02 EST by Prof Sassan Asgari on behalf of School of Biological Sciences