title: Critical role of Casein kinase 2 in hepatitis C NS5A-mediated inhibition of Kv2.1 K(+) channel function.
abstract: Inhibiting injury-induced increases in outward K(+) currents is sufficient to block cell death in cortical neuronal injury models. It is now known that apoptosis is facilitated in hepatocytes by the same K(+) channel as in cortical neurons, namely, the delayed rectifier K(+) channel Kv2.1. The hepatitis C virus (HCV) protein NS5A prevents the apoptosis-enabling loss of intracellular potassium by inhibiting Kv2.1 function and thus blocking hepatocyte cell death. Critically, neurons expressing NS5A1b (from HCV genotype 1b), but not NS5A1a, can be protected from lethal injurious stimuli via a block of Kv2.1-mediated potassium currents. Here, we identify a key component unique to NS5A1b, which is necessary for restricting Kv2.1 currents and establishing neuroprotection. By comparing the sequence differences between NS5A1b and 1a we identify putative casein kinase 2 (CK2) phosphorylation regions unique to the 1b genotype. We show that selective inhibition of CK2 in cortical neurons results in loss of NS5A1b's ability to depress outward potassium currents, and, surprisingly, potentiates currents in non-NS5A-expressing cells. As such, our results suggest that NS5A1b-mediated inhibition of Kv2.1 function is critically dependent on its phosphorylation status at genotypic-specific CK2-directed residues. Importantly, inhibiting NS5A viral replicative function with the novel HCV drug Ledipasvir does not impair the ability of this protein to block Kv2.1 function. This suggests that the modulation of NS5A function by CK2 may be a component of HCV unique to the regulation of apoptosis.
authors.affiliation: Department of Neurobiology and Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Katerina
authors.initials: K
authors.lastname: Clemens
authors.affiliation: Department of Neurobiology and Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: Chung-Yang
authors.initials: CY
authors.lastname: Yeh
authors.affiliation: Department of Neurobiology and Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. Electronic address: redox@pitt.edu.
authors.forename: Elias
authors.initials: E
authors.lastname: Aizenman
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D001562:Benzimidazoles / Q000494:pharmacology; D047390:Casein Kinase II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D005449:Fluorenes / Q000494:pharmacology; D005822:Genetic Vectors; D008297:Male; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation; D017207:Rats, Sprague-Dawley; D051669:Shab Potassium Channels / Q000037:antagonists & inhibitors*; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000998:Antiviral Agents; D001562:Benzimidazoles; D047390:Casein Kinase II; D005449:Fluorenes; D051669:Shab Potassium Channels; D017361:Viral Nonstructural Proteins; C496991:Kcnb1 protein, rat; C586541:ledipasvir; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase