Werner Syndrome (WS) is an autosomal recessive disorder characterized by premature development of aging features. In addition, WS individuals have an increased predisposition to cancers that are mesenchymal in origin. Primary cells derived from WS patients exhibit elevated levels of chromosomal translocations, inversions, and deletions of large segments of DNA and have a high spontaneous mutation rate. The Werner syndrome protein (WRN), mutated in WS, is unique among the RecQ family proteins as it possesses exonuclease and 3? to 5? helicase activities. Thesetwo enzymatic activities have coordinated functions on a variety of structured DNA substrates. Additionally, WRN also has nuclease-independent functions during DNA replication and repair, although these non-enzymatic activities are not well understood. WRN forms dynamic sub-complexes with different factors involved in multiple biological processes.Though accumulating evidence suggests that WRN plays a crucial role in genome stability maintenance pathways, the exact contribution of WRN in preventing genome instability is unclear. Our hypothesis is that WRN suppresses genomic instability by facilitating faithful repair of collapsed replication forks upon replication stress. To accomplish our goals, three specific aims are proposed:
Aim 1. Verify that a single phosphorylation site dictates other post- translational modifications in WRN and its function in genome stability maintenance upon replication stress;
Aim 2. Determine the mechanism used by WRN to stabilize Rad51 at collapsed replication forks;
and Aim 3. Verify that WRN interacts with specific-genomic loci and suppresses mis-joining of collapsed replication forks. Innovative aspects of this proposal include: a. Development of a novel system to identify cell cycle-specific biological functions of WRN. In this system, expression of WRN can be regulated in a cell cycle-specific manner. b. Establishment of a new tool to map genome interaction domains of WRN and precisely identify fidelity of replication-associated DNA double-strand breaks at a single nucleotide level. Results from this genomic sequencing study will provide information on molecular biomarker(s) contributing to the initiation of carcinogenic events in WS. Significantly, deciphering the molecular choreography of WRN, its biochemical activities, post-translational modifications and interaction partners in the fidelity of replication fork processing will provide new insight into the molecular origin of cancer in WS individuals. Our results will ultimately advance our understanding of the pathophysiology of WS.

Public Health Relevance

Werner syndrome (WS) is a rare hereditary disease characterized by premature aging onset and a predisposition to a broad spectrum of rare cancers. The Werner syndrome protein (WRN), defective in WS, plays multiple roles in genome stability maintenance pathways; however, WRN mechanisms involved in carcinogenesis prevention at the nucleotide level are poorly understood. Deciphering the molecular choreography of WRN, its biochemical activities, post-translational modifications and interaction partners in the fidelity of replication fork processing will not only provide new insight into the molecular origin of cancer, but also will advance our understanding of the pathophysiology of WS.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG053341-05
Application #
9901410
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Guo, Max
Project Start
2016-08-01
Project End
2021-03-31
Budget Start
2020-04-01
Budget End
2021-03-31
Support Year
5
Fiscal Year
2020
Total Cost
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390
Mukherjee, Shibani; Sinha, Debapriya; Bhattacharya, Souparno et al. (2018) Werner Syndrome Protein and DNA Replication. Int J Mol Sci 19:
Bhattacharya, Souparno; Asaithamby, Aroumougame (2017) Repurposing DNA repair factors to eradicate tumor cells upon radiotherapy. Transl Cancer Res 6:S822-S839
Bhattacharya, Souparno; Srinivasan, Kalayarasan; Abdisalaam, Salim et al. (2017) RAD51 interconnects between DNA replication, DNA repair and immunity. Nucleic Acids Res 45:4590-4605