Glycogen synthase kinase 3ß participates in late stages of Dengue virus-2 infection

Memórias do Instituto Oswaldo Cruz
Alexandra Milena Cuartas-López, Juan Carlos Gallego-Gómez

Abstract

Viruses can modulate intracellular signalling pathways to complete their infectious cycle. Among these, the PI3K/Akt pathway allows prolonged survival of infected cells that favours viral replication. GSK3β, a protein kinase downstream of PI3K/Akt, gets inactivated upon activation of the PI3K/Akt pathway, and its association with viral infections has been recently established. In this study, the role of GSK3β during Dengue virus-2 (DENV-2) infection was investigated. GSK3β participation in the DENV-2 replication process was evaluated with pharmacological and genetic inhibition during early [0-12 h post-infection (hpi)], late (12-24 hpi), and 24 hpi in Huh7 and Vero cells. We assessed the viral and cellular processes by calculating the viral titre in the supernatants, In-Cell Western, western blotting and fluorescence microscopy. Phosphorylation of GSK3β-Ser9 was observed at the early stages of infection; neither did treatment with small molecule inhibitors nor pre-treatment prior to viral infection of GSK3β reduce viral titres of the supernatant at these time points. However, a decrease in viral titres was observed in cells infected and treated with the inhibitors much later during viral infection. Consistently, the infected ce...Continue Reading

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Methods Mentioned

BETA
Protein Assay
Infrared Imaging
transfection
electrophoresis
fluorescence microscopy
flow cytometry

Software Mentioned

Fiji
Media
Pro Plus
Image
ImageJ

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