Investigation of early events in oncogenic HPV infection

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DESCRIPTION (provided by applicant): The goal of this proposal is to study and compare the early events in the life cycle of oncogenic HPV16, 18, and 31. Specifically we want to identify the precise endocytic pathways used for host cell penetration and identify what types of cellular signalling events are both required for and induced by the process of cell entry. Inherent differences in the cancer biology as well the capsid composition of these oncogenic HPVs warrants the investigation of these viral-host cell interactions. We will determine the kinetics of uptake by both infectivity/neutralization timecourse experiments as well as by direct visualization by confocal and electron microscopy. The specific endocytic routes of entry will be determined through a combination of pharmacological inhibition experiments and direct visualization of colocalization between virions and specific endocytic markers. Signal transduction in response to the endocytic uptake of virions will be assessed by the upregulation of phospho-proteins, the identity of which can be revealed by mass spectroscopy. The effects of pharmacological inhibition of specific kinase and phosphatase activities on HPV intracellular trafficking and infectivity will also be determined. Taken together, results from these studies will help define the molecular events during the initial HPV-host cell interactions and will set the stage for more detailed investigation into specifics of these critical processes (cytosolic translocation, virion disassembly, etc.)