The productive phase of the HPV life cycle is restricted to the uppermost layer of the epithelium, in cells that have normally exited the cell cycle. The E7 protein maintains differentiating cells active in the cell cycle, allowing for productive replication and virion production. Cell cycle deregulation by E7 results in genomic instability that contributes to cancer development. Our long-term goal is to identify mechanisms that regulate the productive phase of the viral life cycle, which is important to understanding how HPV causes cancer. This proposal focuses on determining how HPV commandeers the ATM DNA damage-signaling pathway to facilitate productive replication. We will identify the mechanisms by which HPV activates ATM and determine how HPV utilizes ATM activity to drive viral replication. We hypothesize that HPV highjacks DNA signaling pathways through E7, leading to ATM activation that drives productive replication through homologous recombination repair.
Specific Aims to test this are: (1) To determine how HPV causes DNA damage by determining the contribution of E2F and STAT5 transcription factors to E7-induced ATM signaling. (2) To determine if ATM activity alters viral assembly of chromatin to facilitate DNA repair factor recruitment, as well as the contribution of the histone variant H2AX to viral replication. (3) To determine how DNA double strand break (DSB) repair pathways are regulated in HPV infected cells by using chromosomally integrated reporters for distinct repair pathways. We will also determine the contribution of the ATM target Nbs1, and the homologous recombination repair factors Rad51 and Brca1, to viral replication. Since ATM is essential to the maintenance of genomic stability and the prevention of cancer, it is important to understand how HPV manipulates this pathway to ensure completion of the viral life cycle. These studies will provide insight not only into viral life cycle, but also the potential mechanisms by which HPV induces genomic instability.

Public Health Relevance

A subset of HPV types are the causative agents of cervical cancer, and are also associated several other human cancers. Our studies focus on the interaction between HPV and the DNA damage kinase ATM, a major regulator of genomic stability. Understanding how HPV activates and utilizes ATM signaling for viral replication has the potential to identify targets that could be important clinically for the treatment of HPV- associated diseases.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA181581-02
Application #
8926368
Study Section
Virology - A Study Section (VIRA)
Program Officer
Read-Connole, Elizabeth Lee
Project Start
2014-09-11
Project End
2019-08-31
Budget Start
2015-09-01
Budget End
2016-08-31
Support Year
2
Fiscal Year
2015
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Moody, Cary (2017) Mechanisms by which HPV Induces a Replication Competent Environment in Differentiating Keratinocytes. Viruses 9:
Anacker, Daniel C; Moody, Cary A (2017) Modulation of the DNA damage response during the life cycle of human papillomaviruses. Virus Res 231:41-49
Johnson, Bryan A; Aloor, Heather L; Moody, Cary A (2017) The Rb binding domain of HPV31 E7 is required to maintain high levels of DNA repair factors in infected cells. Virology 500:22-34
Gautam, Dipendra; Moody, Cary A (2016) Impact of the DNA Damage Response on Human Papillomavirus Chromatin. PLoS Pathog 12:e1005613
Chappell, William H; Gautam, Dipendra; Ok, Suzan T et al. (2016) Homologous Recombination Repair Factors Rad51 and BRCA1 Are Necessary for Productive Replication of Human Papillomavirus 31. J Virol 90:2639-52
Anacker, Daniel C; Aloor, Heather L; Shepard, Caitlin N et al. (2016) HPV31 utilizes the ATR-Chk1 pathway to maintain elevated RRM2 levels and a replication-competent environment in differentiating Keratinocytes. Virology 499:383-396
Anacker, Daniel C; Gautam, Dipendra; Gillespie, Kenric A et al. (2014) Productive replication of human papillomavirus 31 requires DNA repair factor Nbs1. J Virol 88:8528-44