JC virus (JCV] is a common human polyomavirus that infects over 70% of the population worldwide. JCV has a restricted cell tropism that is partly due to the initial interaction between the virus and sialic acid containing host cell receptors. We have identified the molecular determinants for interaction between the virus and its cellular receptor, using a combined approach of site-directed mutagenesis and homology-based molecular modeling. Here, we propose incorporating the JCV sialic acid binding domain onto 4-helix bundle proteins, providing a soluble platform for in-depth biophysical characterization of the virus/sialic acid association as well as screening of compounds for potent inhibitors. Preliminary results indicate that the protein platform is a potent inhibitor of JC viral infection and competes with the virus for cell surface receptors. We propose to further characterize the protein template, including comparing the results from mutagenesis studies to those previously carried out for the intact virus. Initial structural studies, employing circular dichroism and highresolution NMR of 15N and 13C enriched samples, display a stable, folded protein and the expected sialic acid binding profile. The structure of the protein template while associated with sialic acid will be determined by NMR, providing the insight for rational design of inhibitors for the association of JCV with its host cell. The JCV templates will also be developed for screening of small compound libraries. Molecules produced from these efforts will allow us to further probe the pathogenesis of JCV induced disease and may lead to novel therapies

Public Health Relevance

The JC virus is the etiological agent for PML, a fatal complication for immunosuppressed individuals, most notably AIDS patients. Here, we propose the characterization of the initial interaction of the JC virus with the host cell employing a model protein system to mimic the JC viral coat, with the aim of developing molecular inhibitors thay may provide a novel route for therapy development

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Program Projects (P01)
Project #
5P01NS065719-05
Application #
8512817
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Project Start
Project End
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
5
Fiscal Year
2013
Total Cost
$258,027
Indirect Cost
Name
Brown University
Department
Type
DUNS #
001785542
City
Providence
State
RI
Country
United States
Zip Code
02912
Maginnis, Melissa S; Nelson, Christian D S; Atwood, Walter J (2015) JC polyomavirus attachment, entry, and trafficking: unlocking the keys to a fatal infection. J Neurovirol 21:601-13
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O'Hara, Bethany A; Rupasinghe, Chamila; Yatawara, Achani et al. (2014) Gallic acid-based small-molecule inhibitors of JC and BK polyomaviral infection. Virus Res 189:280-5
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Neu, Ursula; Allen, Stacy-Ann A; Blaum, Barbel S et al. (2013) A structure-guided mutation in the major capsid protein retargets BK polyomavirus. PLoS Pathog 9:e1003688
Nelson, Christian D S; Carney, Dan W; Derdowski, Aaron et al. (2013) A retrograde trafficking inhibitor of ricin and Shiga-like toxins inhibits infection of cells by human and monkey polyomaviruses. MBio 4:e00729-13
Gee, Gretchen V; O'Hara, Bethany A; Derdowski, Aaron et al. (2013) Pseudovirus mimics cell entry and trafficking of the human polyomavirus JCPyV. Virus Res 178:281-6
Maginnis, Melissa S; Stroh, Luisa J; Gee, Gretchen V et al. (2013) Progressive multifocal leukoencephalopathy-associated mutations in the JC polyomavirus capsid disrupt lactoseries tetrasaccharide c binding. MBio 4:e00247-13
Lipovsky, Alex; Popa, Andreea; Pimienta, Genaro et al. (2013) Genome-wide siRNA screen identifies the retromer as a cellular entry factor for human papillomavirus. Proc Natl Acad Sci U S A 110:7452-7
Yatawara, Achani K; Hodoscek, Milan; Mierke, Dale F (2013) Ligand binding site identification by higher dimension molecular dynamics. J Chem Inf Model 53:674-80

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