Faculty Collaboration Database

Artemisinins target the intermediate filament protein vimentin for human cytomegalovirus inhibition. J Biol Chem 2020 Oct 30;295(44):15013-15028

Date

08/29/2020

Pubmed ID

32855235

Pubmed Central ID

PMC7606667

DOI

10.1074/jbc.RA120.014116

Scopus ID

2-s2.0-85094983683 (requires institutional sign-in at Scopus site)   14 Citations

Abstract

The antimalarial agents artemisinins inhibit cytomegalovirus (CMV) in vitro and in vivo, but their target(s) has been elusive. Using a biotin-labeled artemisinin, we identified the intermediate filament protein vimentin as an artemisinin target, validated by detailed biochemical and biological assays. We provide insights into the dynamic and unique modulation of vimentin, depending on the stage of human CMV (HCMV) replication. In vitro, HCMV entry and viral progeny are reduced in vimentin-deficient fibroblasts, compared with control cells. Similarly, mouse CMV (MCMV) replication in vimentin knockout mice is significantly reduced compared with controls in vivo, confirming the requirement of vimentin for establishment of infection. Early after HCMV infection of human foreskin fibroblasts vimentin level is stable, but as infection proceeds, vimentin is destabilized, concurrent with its phosphorylation and virus-induced calpain activity. Intriguingly, in vimentin-overexpressing cells, HCMV infection is reduced compared with control cells. Binding of artesunate, an artemisinin monomer, to vimentin prevents virus-induced vimentin degradation, decreasing vimentin phosphorylation at Ser-55 and Ser-83 and resisting calpain digestion. In vimentin-deficient fibroblasts, the anti-HCMV activity of artesunate is reduced compared with controls. In summary, an intact and stable vimentin network is important for the initiation of HCMV replication but hinders its completion. Artesunate binding to vimentin early during infection stabilizes it and antagonizes subsequent HCMV-mediated vimentin destabilization, thus suppressing HCMV replication. Our target discovery should enable the identification of vimentin-binding sites and compound moieties for binding.

Author List

Roy S, Kapoor A, Zhu F, Mukhopadhyay R, Ghosh AK, Lee H, Mazzone J, Posner GH, Arav-Boger R

Authors

Ravit Boger MD Chief, Professor in the Pediatrics department at Medical College of Wisconsin
Ayan K. Ghosh PhD Research Scientist I in the Pediatrics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Antiviral Agents
Artemisinins
Binding Sites
Calpain
Cell Cycle
Cells, Cultured
Cytomegalovirus
Cytomegalovirus Infections
Drug Repositioning
Humans
Mass Spectrometry
Phosphorylation
Proteasome Endopeptidase Complex
Vimentin
Virus Replication
Withanolides