Faculty Collaboration Database

NAADP-dependent Ca²⁺ signaling regulates Middle East respiratory syndrome-coronavirus pseudovirus translocation through the endolysosomal system. Cell Calcium 2018 Nov;75:30-41

Date

08/20/2018

Pubmed ID

30121440

Pubmed Central ID

PMC6251489

DOI

10.1016/j.ceca.2018.08.003

Scopus ID

2-s2.0-85051524070 (requires institutional sign-in at Scopus site)   98 Citations

Abstract

Middle East Respiratory Syndrome coronavirus (MERS-CoV) infections are associated with a significant mortality rate, and existing drugs show poor efficacy. Identifying novel targets/pathways required for MERS infectivity is therefore important for developing novel therapeutics. As an enveloped virus, translocation through the endolysosomal system provides one pathway for cellular entry of MERS-CoV. In this context, Ca²⁺-permeable channels within the endolysosomal system regulate both the luminal environment and trafficking events, meriting investigation of their role in regulating processing and trafficking of MERS-CoV. Knockdown of endogenous two-pore channels (TPCs), targets for the Ca²⁺ mobilizing second messenger NAADP, impaired infectivity in a MERS-CoV spike pseudovirus particle translocation assay. This effect was selective as knockdown of the lysosomal cation channel mucolipin-1 (TRPML1) was without effect. Pharmacological inhibition of NAADP-evoked Ca²⁺ release using several bisbenzylisoquinoline alkaloids also blocked MERS pseudovirus translocation. Knockdown of TPC1 (biased endosomally) or TPC2 (biased lysosomally) decreased the activity of furin, a protease which facilitates MERS fusion with cellular membranes. Pharmacological or genetic inhibition of TPC1 activity also inhibited endosomal motility impairing pseudovirus progression through the endolysosomal system. Overall, these data support a selective, spatially autonomous role for TPCs within acidic organelles to support MERS-CoV translocation.

Author List

Gunaratne GS, Yang Y, Li F, Walseth TF, Marchant JS

Author

Jonathan S. Marchant PhD Chair, Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin

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

Benzylisoquinolines
Calcium Signaling
Cell Line
Endosomes
Furin
Gene Knockdown Techniques
Humans
Ion Channels
Lysosomes
Middle East Respiratory Syndrome Coronavirus
NADP
Reproducibility of Results
Spike Glycoprotein, Coronavirus
Voltage-Gated Sodium Channels