Download Title: Intracellular Calcium Regulation in JC Polyomavirus Infection

Title: Intracellular Calcium Regulation in JC Polyomavirus Infection
Authors: Ashley N. Soucy1,2, Jeanne K. DuShane2, Melissa S. Maginnis2
Affiliations: 1Honors College, 2Department of Molecular and Biomedical Sciences, University of
Maine, Orono ME
Background and Objectives: The majority of the human population is infected with JC
polyomavirus (JCPyV). The virus establishes a persistent, asymptomatic infection in the kidney
of healthy individuals. In immunosuppressed individuals, JCPyV can migrate to the CNS and
cause the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML).
Previous studies suggested that calcium (Ca2+) signaling is necessary for the activation of
transcription factors required for JCPyV infection. The goal of this study is to define how
intracellular calcium flux influences the infectious life cycle and pathogenicity of the virus.
Methods: Glial cells were pretreated with 2-APB, an antagonist of IP3-mediated Ca2+ release
from the ER, or U73122, an inhibitor of PLC to prevent hydrolysis of PIP2 to IP3, then infected
with JCPyV. Infection was quantified by indirect immunofluorescence microscopy. Cells treated
with 2-APB were tested for the capacity to support virus binding by flow cytometry.
Results: 2-APB inhibited JCPyV infection but had no effect on viral attachment, while U73122
resulted in reduced infection.
Discussion and Conclusions: Inhibition of IP3-mediated Ca2+ release from the ER inhibits
JCPyV infection in a dose-dependent manner, yet this inhibition had no effect on viral
attachment. Inhibition of PLC, also reduces JCPyV infection. Taken together, these data
suggest that Ca2+ release from the ER is required for JCPyV infection at a post-attachment step.
Grant Support: Research reported in this project was supported by an Institutional
Development Award (IDeA) from the National Institute of General Medical Sciences of the
National Institutes of Health under grant number P20GM103423, a Frederick H. Radke
Undergraduate Research Fellowship, and The University of Maine.