These studies will continue a long-standing collaboration between laboratories at Cornell University and Purdue University which involve genetic, virological, and structural studies of parvovirus capsids, their cellular receptors, and the antibodies that neutralize them. Parvoviruses cause widespread and serious disease in humans and other animals, and are genetically and structurally simple. However, they display the behaviors that are seen for most animal viruses, including receptor binding, endosomal uptake and cell infection, cytoplasmic and nuclear trafficking, and efficient neutralization by antibodies. In most of the work proposed here we are studying canine parvovirus (CPV) and feline panleukopenia virus (FPV), which are >99.5% identical in DNA sequence, yet which differ in host range through mutations in the capsid protein which affect specific receptor binding. In our recent work we have defined capsid structural variation, capsid assembly processes, antibody-capsid interactions, and identified the transferrin receptor (TfR) as a major receptor for cell binding and infection. We will continue studies in 5 areas. (1) Mutagenesis of the feline TfR and the capsid to define the interactions between those molecules that control viral binding and allow infection; (2) Use cryo-electron microscopy and X-ray crystallography to define in detail the structures of the interacting sites on the feline and canine TfRs and the capsid; (3) Analyze structural variation of the capsid, and define changes induced by TfR binding leading to infection of cells, as well as those induced by antibody binding leading to neutralization. Antibody domains expressed on cells as fusions with the TfR will be used as alternative receptors to determine whether they can mediate virus infection of cells; (4) Examine the role of a pore in the capsid structure and determine whether it controls exposure of the N-termini of the VP1 or VP2 proteins, or the 3' or 5' ends of the viral DNA. The infectivity and release of sequences from the mutant capsids will be examined and compared to wild type viruses; (5) Determine the high resolution structure of the human parvovirus B19 capsid and correlate that structure with the known properties of the virus.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI033468-13
Application #
6855118
Study Section
Virology Study Section (VR)
Program Officer
Park, Eun-Chung
Project Start
1992-12-01
Project End
2008-01-31
Budget Start
2005-02-01
Budget End
2006-01-31
Support Year
13
Fiscal Year
2005
Total Cost
$319,904
Indirect Cost
Name
Cornell University
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
872612445
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Lyi, Sangbom Michael; Tan, Min Jie Alvin; Parrish, Colin R (2014) Parvovirus particles and movement in the cellular cytoplasm and effects of the cytoskeleton. Virology 456-457:342-52
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Kaufmann, Barbel; El-Far, Mohamed; Plevka, Pavel et al. (2011) Structure of Bombyx mori densovirus 1, a silkworm pathogen. J Virol 85:4691-7
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Nelson, Christian D S; Palermo, Laura M; Hafenstein, Susan L et al. (2007) Different mechanisms of antibody-mediated neutralization of parvoviruses revealed using the Fab fragments of monoclonal antibodies. Virology 361:283-93

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