1. LA PÉRDIDA DE FUNCIÓN DE LA QUINASA DEPENDIENTE DE CICLINA 5 (CDK5) ALTERA EL CITOESQUELETO Y REDUCE LA INFECCIÓN in vitro POR EL VIRUS DEL DENGUE 2.
- Author
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ROA-LINARES, Vicky C. and GALLEGO-GÓMEZ, Juan Carlos
- Subjects
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VIRAL envelope proteins , *GENE silencing , *DENGUE viruses , *VIRAL genomes , *EPITHELIAL cells , *MICROTUBULES , *CYTOSKELETON - Abstract
Cyclin-Dependent Kinase 5 (CDK5) regulates several functions in neurons, endothelial, and epithelial cells, including the cytoskeleton dynamics. Likewise, it has been reported that some cytoskeleton elements, such as actin filaments and microtubules, play an essential role during Dengue virus (DENV) infection. This work aimed to evaluate the role of CDK5 during DENV-2 infection by two methods, chemical inhibition, and gene silencing. The antiviral activity of roscovitine was evaluated using Plaque Forming Units (PFU) assay. The transfection efficiency and knockdown of CDK5, using artificial miRNAs, was carried out by flow cytometry. The effect on the viral envelope protein and cytoskeleton elements was evidenced by advanced fluorescence microscopy and image analysis. Roscovitine showed antiviral activity in pre and post-infection stages in a dose-dependent manner. Treatment with roscovitine and miRCDK5 decrease the amount of CDK5 in uninfected cells. In cells infected and transfected with miRCDK5, as well as treated with the inhibitor, a significant reduction of the viral envelope protein was observed; however, no significant reduction of CDK5 was found. Also, evident morphological cellular changes were observed during the treatment with roscovitine in infected cells. The results indicate the potential participation of CDK5 during DENV-2 infection, possibly mediating protein translation or replication of the viral genome through the cytoskeletal dynamics regulation. Additional data are required to clarify the mechanistic of these phenomena using alternative methods [ABSTRACT FROM AUTHOR]
- Published
- 2019
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