REGULATION OF HUMAN CYTOMEGALOVIRUS MAJOR IMMEDIATE EARLY GENE EXPRESSION DURING LYTIC REPLICATION

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  • March 22, 2019
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  • Arend, Kyle
    • Affiliation: School of Medicine, Department of Microbiology and Immunology
Abstract
  • Human cytomegalovirus (HCMV) is a significant cause of disease in immune-compromised adults and immune naïve newborns. No vaccine exists to prevent HCMV infection, and current antiviral therapies have toxic side effects that limit the duration and intensity of their use. Expression of the HCMV major immediate early (MIE) proteins, IE1 and IE2, is critical for the establishment of lytic infection and reactivation from viral latency. Defining the mechanisms controlling IE1 and IE2 expression is therefore important for understanding how HCMV regulates its replicative cycle. In Chapter 2 we identified several novel transcripts encoding full-length IE1 and IE2 proteins during HCMV lytic replication. While the canonical MIE mRNA was the most abundant transcript at immediate early times, the novel MIE transcripts accumulated to equivalent levels as the known MIE transcript later in infection and were found associated with polyribosomes. These results expand our understanding of the sequences controlling IE1 and IE2 expression by defining novel transcriptional units controlling the expression of full-length IE1 and IE2 proteins. Beyond transcriptional regulation, relatively little is known about the post-transcriptional mechanisms that control IE1 and IE2 protein synthesis. In Chapter 3 we found that the canonical MIE 5’ untranslated region (5' UTR) has a positive role in translation control of reporter genes during transfection. We also found that the MIE 5’UTR was necessary for efficient IE1 and IE2 mRNA translation as well as viral replication during infection. These results demonstrate that the shared 5’ UTR of the IE1 and IE2 mRNA is a critical determinant of efficient HCMV replication. Virus-induced changes in infected cells are often driven by changes in cellular kinase activity. In Chapter 4 we applied a kinase capture technique, MIB-MS kinome profiling, to quantitatively measure perturbations in >240 cellular kinases simultaneously in cells infected with HCMV. Based on the kinome data, we identified three compounds currently being studied in clinical trials that inhibited HCMV replication. These results show the utility of MIB-MS kinome profiling for identifying existing kinase inhibitors that can potentially be repurposed as novel antiviral drugs to limit the time and cost of new antiviral drug development.
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Advisor
  • Heise, Mark
  • Wolfgang, Matthew
  • Lemon, Stanley
  • Moorman, Nathaniel
  • Moody, Cary
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2018
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