Ionizing Radiation kills cancer cells by generating DNA damage. Resistance to Ionizing Radiation is generally determined by DNA repair mechanisms that can repair radiation-induced DNA damage. The most toxic effect of radiation is represented by the formation of double stranded DNA breaks. These breaks can be repaired by Homologous Recombination, and Non-Homologous End Joining DNA repair mechanisms. Inhibition of these mechanisms results in increased radiosensitivity since repair of radiation-induced double strand breaks is blocked. Thus, understanding the molecular mechanisms of double strand break repair is essential for designing novel radiosensitization therapies. We have recently uncovered a novel mechanism that regulates radioresistance, centered on the protein PARP14. PARP14 is a mono-ADP-ribosyltransferase, which unlike its well-known cousin PARP1 is unable to catalyze poly-ADP-ribose chain formation but can only transfer a single molecule of ADP-ribose to substrates. Mono-ADP-ribosylation is a still mysterious post-translational modification, and its functions in the cell are poorly characterized. Or preliminary results presented here show that PARP14 promotes repair of double strand breaks by activating Homologous Recombination DNA repair. Moreover, our preliminary results presented here indicate a mechanism for this activation: PARP14 mono-ADP-ribosylates the recombination factor RAD51 to promote RAD51 removal from D-loops thus allowing D-loop extension past the break and complete the recombination reaction. Thus, our work shows that PARP14 inhibition results in radiosensitivity by downregulating Homologous Recombination-dependent repair of radiation-induced double strand DNA breaks. In this proposal, we test the hypothesis that PARP14 promotes the repair of radiation-induced double strand DNA breaks by Homologous Recombination, through RAD51 mono-ADP-ribosylation. We will employ a comprehensive, integrative approach to study: 1) the impact of PARP14 on the repair of radiation-induced DNA damage by Homologous Recombination and other DNA repair mechanisms, and 2) the role of RAD51 mono- ADP-ribosylation by PARP14 in repair of radiation-induced double strand breaks. Our study is highly innovative since it addresses a novel, previously unrecognized function of mono- ADP-ribosylation in radioresistance. Our work will unravel how mono-ADP-ribosylation by PARP14 promotes radiation resistance, and describe novel radiosensitization approaches, based on PARP14 inhibition.

Public Health Relevance

Radiation therapy is one of the main cancer treatments available. However, some tumors are resistant to radiation resulting in cancer recurrence. Radiation kills cancer cells by attacking their DNA. Understanding how cancer cells repair these radiation-induced lesions, will allow the development of new drugs that can sensitize tumors to radiation. Here we investigate a new mechanism for DNA damage repair we recently discovered.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES026184-03
Application #
9420616
Study Section
Radiation Therapeutics and Biology Study Section (RTB)
Program Officer
Reinlib, Leslie J
Project Start
2016-02-01
Project End
2021-01-31
Budget Start
2018-02-01
Budget End
2019-01-31
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Pennsylvania State University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
129348186
City
Hershey
State
PA
Country
United States
Zip Code
17033
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Choe, Katherine N; Nicolae, Claudia M; Constantin, Daniel et al. (2016) HUWE1 interacts with PCNA to alleviate replication stress. EMBO Rep 17:874-86