Human cytomegalovirus (HCMV) is a ubiquitous and significant pathogen that stimulates cell cycle progression. HCMV is a cofactor for cardiovascular diseases with significant proliferative components such as coronary restenosis and transplant vascular sclerosis (TVS), and may play a role in certain types of cancers, most notably, malignant gliomas (GBMs). Cellular proliferation and transformation is regulated by the retinoblastoma family of tumor suppressors (Rb, p107, and p130). The Rb proteins, in conjunction with the E2F family of transcription factors, regulate the expression of cellular genes required for DNA replication such as nucleotide biosynthetic enzymes (NBEs) of the de novo pathway, as well as secreted factors involved in wound healing and angiogenesis. The expression of E2F-responsive genes is inhibited when the hypophosphorylated forms of the Rb proteins are bound to the E2F proteins. Degradation of the hypophosphorylated Rb proteins, or their hyperphosphorylation disrupts their complexes with E2F, thus allowing free E2F to activate gene expression. HCMV encodes four proteins that modulate the Rb-E2F pathway: pp71, IE1, IE2, and the focus of this application, UL97. Because HCMV is associated with proliferative diseases and encodes multiple proteins that modulate the Rb-E2F pathway, we hypothesize that the Rb-E2F pathway plays a critical role in HCMV replication and pathogenesis. Here we propose to explore the roles of the Rb, E2F, and UL97 proteins during HCMV infection. UL97 is a member of a family of conserved herpesvirus protein kinases (CHPKs) that directly phosphorylates the Rb protein and inactivates it, leading to the induction of cellular E2F-responsive genes. We hypothesize that some of these E2F-responsive gene products (the NBEs) are responsible for the UL97-mediated stimulation of viral DNA replication. We further speculate that other E2F-responsive gene products induced by UL97 (secreted wound healing and angiogenesis factors) may in turn be partially responsible for proliferative pathologies associated with HCMV infection, such as restenosis, TVS, and GBMs. The long-term goal of this project is to determine how Rb inactivation by UL97 and other HCMV proteins impacts cell cycle progression, HCMV replication, and viral pathogenesis in both cell autonomous and non-cell-autonomous manners, and to use this information to design therapies to treat proliferative diseases associated with HCMV infection. PHS 398/2590 (Rev. 05/01) Page 1 Continuation Format Page

Public Health Relevance

Some viruses cause cancer, a disease of uncontrolled cellular growth. Human cytomegalovirus (HCMV) infects almost everyone, causes severe disease especially in newborn infants and transplant patients, and may play a role in many different types of cancer. We are studying how HCMV causes cells to grow in the hopes of finding drugs to stop that process, and that could be used as anti-viral and anti-cancer treatments.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI080675-05
Application #
8651405
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Beisel, Christopher E
Project Start
2010-05-01
Project End
2015-04-30
Budget Start
2014-05-01
Budget End
2015-04-30
Support Year
5
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Public Health & Prev Medicine
Type
Graduate Schools
DUNS #
City
Madison
State
WI
Country
United States
Zip Code
53715
Umaña, Angie C; Iwahori, Satoko; Kalejta, Robert F (2018) Direct Substrate Identification with an Analog Sensitive (AS) Viral Cyclin-Dependent Kinase (v-Cdk). ACS Chem Biol 13:189-199
Liu, Che; Clark, Paul A; Kuo, John S et al. (2017) Human Cytomegalovirus-Infected Glioblastoma Cells Display Stem Cell-Like Phenotypes. mSphere 2:
Iwahori, Satoko; Umaña, Angie C; VanDeusen, Halena R et al. (2017) Human cytomegalovirus-encoded viral cyclin-dependent kinase (v-CDK) UL97 phosphorylates and inactivates the retinoblastoma protein-related p107 and p130 proteins. J Biol Chem 292:6583-6599
Iwahori, Satoko; Kalejta, Robert F (2017) Phosphorylation of transcriptional regulators in the retinoblastoma protein pathway by UL97, the viral cyclin-dependent kinase encoded by human cytomegalovirus. Virology 512:95-103
Kuny, Chad V; Kalejta, Robert F (2016) Human Cytomegalovirus Can Procure Deoxyribonucleotides for Viral DNA Replication in the Absence of Retinoblastoma Protein Phosphorylation. J Virol 90:8634-43
Iwahori, Satoko; Hakki, Morgan; Chou, Sunwen et al. (2015) Molecular Determinants for the Inactivation of the Retinoblastoma Tumor Suppressor by the Viral Cyclin-dependent Kinase UL97. J Biol Chem 290:19666-80
VanDeusen, Halena R; Kalejta, Robert F (2015) Deficiencies in Cellular Processes Modulated by the Retinoblastoma Protein Do Not Account for Reduced Human Cytomegalovirus Replication in Its Absence. J Virol 89:11965-74
Sun, Xiaoping; Bristol, Jillian A; Iwahori, Satoko et al. (2013) Hsp90 inhibitor 17-DMAG decreases expression of conserved herpesvirus protein kinases and reduces virus production in Epstein-Barr virus-infected cells. J Virol 87:10126-38
Ranganathan, Padhma; Clark, Paul A; Kuo, John S et al. (2012) Significant association of multiple human cytomegalovirus genomic Loci with glioblastoma multiforme samples. J Virol 86:854-64
Dziurzynski, Kristine; Chang, Susan M; Heimberger, Amy B et al. (2012) Consensus on the role of human cytomegalovirus in glioblastoma. Neuro Oncol 14:246-55

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