Cellular microRNA-miR-548g-3p modulates the replication of dengue virus - 08/05/15

Doi : 10.1016/j.jinf.2014.12.001

Weitao Wen ^a,^b,^c,^{^{h
These authors contributed equally to this work.}}, Zhenjian He ^a,^b,^c,^d,^{^{h
These authors contributed equally to this work.}}, Qinlong Jing ^d,^e,^{^{h
These authors contributed equally to this work.}}, Yiwen Hu ^a,^b,^c, Cuiji Lin ^a,^b,^c, Rui Zhou ^f, Xiaoqun Wang ^f, Yangfan Su ^f, Jiehao Yuan ^f, Zhenxin Chen ^f, Jie Yuan ^a,^b,^g, Jueheng Wu ^a,^b,^c, Jun Li ^a,^b,^g, Xun Zhu ^a,^b,^c,^⁎ , Mengfeng Li ^a,^b,^c,^⁎
^a Key Laboratory of Tropical Disease Control (Sun Yat-sen University), Ministry of Education, Guangzhou 510080, China
^b Guangdong Province Key Laboratory of Functional Molecules in Oceanic Microorganism (Sun Yat-sen University), Bureau of Education, Guangzhou 510080, China
^c Department of Microbiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China
^d Department of Medical Statistics and Epidemiology, School of Public Health, Sun Yat-Sen University, Guangzhou 510080, China
^e Department of Infectious Diseases, Guangzhou Center for Disease Control and Prevention, Guangzhou 510440, China
^f Department of Clinical Medicine, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China
^g Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China

^∗Corresponding author. Zhongshan School of Medicine, Sun Yat-sen University, 74 Zhongshan Road II, Guangzhou, Guangdong 510080, China. Tel.: +86 20 87335868; fax: +86 20 87330209.^∗∗Corresponding author. Zhongshan School of Medicine, Sun Yat-sen University, 74 Zhongshan Road II, Guangzhou, Guangdong 510080, China. Tel.: +86 20 87332748; fax: +86 20 87330209.

Summary

Objective

It has been well recognized that microRNA plays a role in the host–pathogen interaction network. The significance of microRNA in the regulation of dengue virus (DENV) replication, however, remains unknown. The objective of our study was to determine the biological function of miR-548g-3p in modulating the replication of dengue virus.

Methods

Here we report that employment of a microRNA target search algorithm to analyze the 5′ untranslated region (5′UTR) consensus sequences of DENV (DENV serotypes 1–4) led to a discovery that miR-548g-3p directly targets the stem loop A promoter element within the 5′UTR, a region essential for DENV replication. Real-time PCR was used to measure the expression levels of miR-548g-3p under DENV infection. We performed overexpression and inhibition assays to test the role of miR-548g-3p on DENV replication. The protein and mRNA levels of interferon were measured by ELISA and real-time PCR respectively.

Result

We found that overexpression of miR-548g-3p suppressed multiplication of DENV 1, 2, 3 and 4, and that miR-548g-3p was also found to interfere with DENV translation, thereby suppressing the expression of viral proteins.

Conclusion

Our results suggest that miR-548g-3p directly regulates DENV replication and warrant further study to investigate the feasibility of microRNA-based anti-DENV approaches.

Le texte complet de cet article est disponible en PDF.

Highlights

•	miR-548g-3p directly targets the SLA promoter element within the 5′UTR of DENV.
•	Overexpression of miR-548g-3p suppressed replication of DENV.
•	The anti-viral effect of miR-548g-3p is independent of interferon signaling.
•	The anti-viral effect of miR-548g-3p is not due to cell cycle arrest.

Le texte complet de cet article est disponible en PDF.

Keywords : Dengue virus, microRNA, miR-548g-3p, Viral replication

Plan

Introduction

Materials and methods

Computational analysis

Cell culture

Virus

Plasmid construction and transfection

Cell viability assay

Luciferase reporter assays

Western blotting analysis

Results

5′UTR of DENV genome contains potential miRNA binding sites

Direct targeting of the 5′UTR of DENV2 genome by miR-548g-3p

miR-548g-3p overexpression suppresses the replication of DENV

miR-548g-3p overexpression delays the cytopathic effect of DENV2

Silencing miR-548g-3p promotes virus replication

The anti-viral effect of miR-548g-3p is independent of interferon signaling

The anti-viral effect of miR-548g-3p is not due to cell cycle arrest

Discussion

Export

Vol 70 - N° 6

P. 631-640 - juin 2015 Retour au numéro

Article précédent

High hepatitis E virus seroprevalence with absence of chronic infection in HIV-infected patients
Antonio Rivero-Juarez, Loreto Martinez-Dueñas, Antonio Martinez-Peinado, Angela Camacho, Celia Cifuentes, Ana Gordon, Mario Frias, Julian Torre-Cisneros, Juan A. Pineda, Antonio Rivero

| Article suivant

Genotype shift in human coronavirus OC43 and emergence of a novel genotype by natural recombination
Yue Zhang, Jianguo Li, Yan Xiao, Jing Zhang, Ying Wang, Lan Chen, Gláucia Paranhos-Baccalà, Lili Ren, Jianwei Wang

Bienvenue sur EM-consulte, la référence des professionnels de santé.
L’accès au texte intégral de cet article nécessite un abonnement.

Déjà abonné à cette revue ?

connectez-vous ou créez un compte

Cellular microRNA-miR-548g-3p modulates the replication of dengue virus - 08/05/15

Summary

Objective

Methods

Result

Conclusion

Highlights

Plan

Export citations

Fichier

Contenu

Accès rapides

Mon compte

Aide & support

Plateformes Elsevier Masson

Déclaration CNIL