Characterization of BKPyV Interaction with Disialylated Gangliosides and PKC Regulated Viral Entry

Allen, Stacy-ann A.

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Year:: 2013
Contributor:: Allen, Stacy-ann A (creator); Atwood, Walter (Director); Reichner, Johnathan (Reader); Page, Rebecca (Reader); Shank, Peter (Reader); DiMaio, Daniel (Reader); Brown University. BIOMED: Pathobiology (sponsor)
Genre:: theses
Subject:: receptor; signalling; Viruses
Extent:: xvii, 150 p.
DOI: https://doi.org/10.7301/Z0F18X2J

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Abstract:: The human polyomavirus BK is a clinically relevant virus that causes the diseases Polyomavirus Induced Nephropathy and Hemorrhagic Cystitis in immune suppressed kidney and bone marrow transplant patients. BKPyV attachment to receptors on host cells has implications for virus spread, tissue and species tropism. As such, the interaction between BKPyV and host cellular receptors and the signaling events that facilitate entry are significant areas of interest in the field. Gangliosides GD1b and GT1b are receptors for BKV. A common shared feature is the presence of a disialic acid moiety. Using viral infection assays we show that in addition to GD1b and GT1b, the disialylated gangliosides GD3 and GD2 can serve as receptors for BKPyV. NMR spectroscopy identifies the α2,8-disialic acid motif as the primary binding epitope for BKPyV. The crystal structure of the BKPyV capsid protein VP1 in complex with GD3 reveals contacts with both sialic acid moieties, providing a basis for the observed specificity. A structure-based model of the GD1b interaction suggests additional, affinity-enhancing contacts, which when evaluated, subtly influence BKPyV infection. Comparison of the receptor binding surfaces of BKPyV and SV40 reveals an amino acid at position 68 in BKPyV VP1 as the major determinant of specificity. Mutation of this amino acid from lysine in BKPyV to serine in SV40 switches the specificity of BKPyV to recognize GM1 versus GD3. Following interaction with its receptors, entry of BKPyV is mediated by caveolae endocytosis which is regulated by Protein Kinase C (PKC). We investigate the role of PKC in BKPyV infection using the PKC inhibitor Bisindolylmaleimide I. Transient inhibition of PKC, which exploits the reversibility of the inhibitor, results in virus susceptibility to BKPyV antibody neutralization and an increase in membrane localization of PKC. Removal of inhibition increases infection. Additionally, exogenous addition of the BKPyV ganglioside receptor GT1b stimulates increased endocytosis of BKPyV and in combination with PKC inhibition, causes a synergistic increase in infection. The data demonstrates that the disialylated gangliosides GD3, GD2, GD1b and GT1b are important for BKPyV infection and that glycosphingolipid mediated caveolae uptake of BKPyV is regulated by PKC.
Notes:: Thesis (Ph.D. -- Brown University (2013)

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Allen, Stacy-ann A., "Characterization of BKPyV Interaction with Disialylated Gangliosides and PKC Regulated Viral Entry" (2013). Pathobiology Theses and Dissertations. Brown Digital Repository. Brown University Library. https://doi.org/10.7301/Z0F18X2J

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Pathobiology Theses and Dissertations

Theses and Dissertations for the Pathobiology department.
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