Identification of potassium and calcium channel inhibitors as modulators of polyomavirus endosomal trafficking
Autor: | Dobson, SJ, Mankouri, J, Whitehouse, A |
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Rok vydání: | 2020 |
Předmět: |
0301 basic medicine
verapamil Endosome viruses 030106 microbiology Endosomes Simian virus 40 tetrandrine Benzylisoquinolines two pore channel Virus Article Cell Line 03 medical and health sciences Viral entry Cell Movement Virology Chlorocebus aethiops Drug Discovery Potassium Channel Blockers Animals Humans Vero Cells Pharmacology biology Chemistry Endoplasmic reticulum Calcium channel endosomal fusion ion channels Virus Internalization biology.organism_classification Calcium Channel Blockers Polyomaviridae Cell biology 030104 developmental biology Two-pore channel HEK293 Cells Merkel cell polyomavirus Nuclear transport Polyomavirus |
Zdroj: | Antiviral Research |
ISSN: | 1872-9096 0166-3542 |
Popis: | During virus entry, members of the Polyomaviridae transit the endolysosomal network en route to the endoplasmic reticulum (ER), from which degraded capsids escape into the cytoplasm and enter the nucleus. Emerging evidence suggests that viruses require both endosomal acidification and the correct ionic balance of K+ and Ca2+ ions in endosomes for correct virus trafficking and genome release. Here, using two polyomaviruses with different capsid architectures, namely Simian virus 40 (SV40) and Merkel cell polyomavirus (MCPyV), we describe methods to rapidly quantify virus infection using IncuCyte ZOOM imaging analysis, and use this system to investigate the role of both K+ and Ca2+ channels during the early stages of virus entry. Using broad spectrum blockers of both K+ and Ca2+ channels to specifically target host cell ion channel functionality, we show that MCPyV, but not SV40 can be inhibited by K+ channel modulators, whilst both viruses are restricted by the broad spectrum Ca2+ channel inhibitor verapamil. Using a panel of more specific Ca2+ blockers, we show that both MCPyV and SV40 are dependent on the activity of two-pore Ca2+ channels (TPCs), as the TPC-specific blocker tetrandrine could prevent the capsid disassembly and nuclear transport required for virus entry. We therefore reveal a novel target to restrict the entry of polyomaviruses, which given the known role of TPCs during endolysosomal-ER fusion, is likely to be applicable to other viruses that transit this pathway. Highlights • We describe novel high-throughput assays to study SV40 and MCPyV infection. • MCPyV, but not SV40, is sensitive to K+ channel inhibition. • Verapamil inhibits MCPyV and SV40 infection. • Tetrandrine is a potent inhibitor of MCPyV and SV40 infection. • Two-pore channel 1/2 activity is essential for polyomavirus entry. |
Databáze: | OpenAIRE |
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