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Access Type

WSU Access

Date of Award

January 2024

Degree Type

Dissertation

Degree Name

Ph.D.

Department

Immunology and Microbiology

First Advisor

Philip E. Pellett

Abstract

Human cytomegalovirus (HCMV) induces changes to most components of the host-cell environment during infection, including the proteome, secretome, and interactome. There are host-host, host-virus, and virus-virus interactions required for productive replication and release of virions: the infectious particles. Our previous systems-level analysis suggests that virion release is dependent on cellular regulators of exosome biogenesis and secretion: Rab 3 and 27. Here, we assess the complete spatial and temporal proteome of Rab proteins to identify their presence and potential roles in infected cells, virions, infected cell exosomes, and host-virus interactions. To further characterize the potential functions of the identified host-virus interactions, we developed a method for integrating multiple types of omic data: Multi-Dimensional Viromics. We used this method to i) create an atlas integrating temporal mRNA and protein expression, intravirion and cellular localization, interacting partners, and virus replication dependence for 129 HCMV proteins, ii) implement temporal normalization of protein abundances to highlight early-stage infection interactions, and iii) perform Gene Ontology and spatial/temporal proteomic analysis for 3400+ host-virus interacting proteins. Using this tool, we can now readily identify knowledge gaps and previously underappreciated potential host-virus interactions, and we provide a framework for integrated omic analyses for other biological models. We then used our multi-dimensional analysis method to characterize the novel interactions between Rab 3 and 27 with HCMV protein US27. Consistent with previous results, our bioinformatic analysis suggests that US27 can act as an activator of these secretory Rab proteins and is important for virion egress. This systems-level analysis tool can facilitate hypothesis-driven experimental design and help us better understand and appreciate the dynamics of host-virus biology. Collectively, this work contributes to our understanding of host and virus protein interactions required for HCMV infection and showcases novel experimental and bioinformatic systems to study host-virus interactions.

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