Cyclin-Dependent Kinases 8 and 19 Regulate Host Cell Metabolism during Dengue Virus Serotype 2 Infection.

Molly Butler,James E Dilisio,David F Ackart,Connie D Brewster,Rushika Perera,Sandra L Quackenbush,Nunya Chotiwan,Brendan K Podell,Joel Rovnak,Randall J Basaraba

doi:10.3390/v12060654

Molly Butler, James E Dilisio + Show 8 more

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https://doi.org/10.3390/v12060654

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Journal: Viruses	Publication Date: Jun 17, 2020
Citations: 8	License type: CC BY 4.0

Affiliation: Colorado State University

Abstract

Dengue virus infection is associated with the upregulation of metabolic pathways within infected cells. This effect is common to infection by a broad array of viruses. These metabolic changes, including increased glucose metabolism, oxidative phosphorylation and autophagy, support the demands of viral genome replication and infectious particle formation. The mechanisms by which these changes occur are known to be, in part, directed by viral nonstructural proteins that contact and control cellular structures and metabolic enzymes. We investigated the roles of host proteins with overarching control of metabolic processes, the transcriptional regulators, cyclin-dependent kinase 8 (CDK8) and its paralog, CDK19, as mediators of virally induced metabolic changes. Here, we show that expression of CDK8, but not CDK19, is increased during dengue virus infection in Huh7 human hepatocellular carcinoma cells, although both are required for efficient viral replication. Chemical inhibition of CDK8 and CDK19 with Senexin A during infection blocks virus-induced expression of select metabolic and autophagic genes, hexokinase 2 (HK2) and microtubule-associated protein 1 light chain 3 (LC3), and reduces viral genome replication and infectious particle production. The results further define the dependence of virus replication on increased metabolic capacity in target cells and identify CDK8 and CDK19 as master regulators of key metabolic genes. The common inhibition of CDK8 and CDK19 offers a host-directed therapeutic intervention that is unlikely to be overcome by viral evolution.

Highlights

According to the World Health Organization, dengue virus (DENV) is endemic in over 100 countries throughout tropical and subtropical regions worldwide
We saw the sharpest increase in cyclin-dependent kinase 8 (CDK8) mRNA expression between 24 and 48 hpi in DENV2-infected cells relative to mock-infected cells and cells treated with UV-treated DENV2 (UV)-inactivated virus (Figure 1B), and subsequently observed a 2.3-fold increase in CDK8 mRNA levels at 36 hpi in six biological replicates (Figure 1C; mean = 2.30 +/− 0.09, p < 0.0001)
As with hexokinase 2 (HK2) (Figure 4), We found that the level of light chain 3 (LC3) mRNA increased over the course of DENV2 infection coincident the viral induction of LC3 mRNA expression was significantly reduced in cells treated with Senexin with the increases in CDK8 and HK2 (Figures 1, 4 and 5A)

Summary

Introduction

According to the World Health Organization, dengue virus (DENV) is endemic in over 100 countries throughout tropical and subtropical regions worldwide. They estimate that 500,000 people with severe dengue require hospitalization each year, with an estimated 2.5% fatality rate [1]. Viral replication places increased demands for production of ATP and metabolites on the host cell [3,4,5]. To meet these demands, viruses manipulate the intracellular environment of the host cell by remodeling cellular structures and by reprogramming cellular metabolism to increase the expression of key metabolic enzymes [6,7,8,9,10,11,12,13,14]

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