Cancer cells possess two properties which place an extreme burden on the DNA replication and repair machinery: they divide rapidly and are often polyploid. These characteristics may require novel strategies for DNA replication that are not utilized during normal cell division. As an example, the expression of DNA polymerase theta, an error-prone translesion polymerase, is frequently upregulated in cancer cells and this overexpression correlates with excessive chromosomal damage and a negative clinical outcome. We have recently found that Drosophila lacking polymerase theta have abnormal phenotypes and display genome instability in tissues that are characterized by rapid S phases and/or polyploidy, including early stage embryos, follicle cells in the female germline, and histoblasts. In addition, we have identified roles for translesion polymerases in homologous recombination repair These preliminary data establish Drosophila as an excellent system in which to investigate tissue- and cellspecific functions of translesion polymerases and provide an opportunity to test the hypothesis that translesion polymerases play important roles in cells that experience endogenous replication stress. We will utilize a novel lacZ reporter system to assess the frequency and types of mutations that arise during replication and homologous recombination repair when polymerase theta and other translesion polymerases are mutated or overexpressed. In addition, we will collaborate with the Lovett and Freudenreich labs to test whether trinucleotide repeats and sequences that form quasi-palindromes experience heightened instability when located near double-strand breaks or under conditions of endogenous replication stress. Together, these studies will significantly advance our long-term goal to understand how the use and misuse of translesion polymerases contributes to genome instability in cancer cells.

Public Health Relevance

The activity of error-prone, translesion DNA polymerases is carefully regulated to prevent the accumulation of mutations that can lead to the development and progression of cancer. This project will characterize how translesion polymerases are utilized and controlled in a model metazoan during specific periods of development and cell division. Through these studies, we will gain an improved understanding of how dysregulation of translesion polymerases can lead to genomic instability characteristic of cancer cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
1P01GM105473-01A1
Application #
8666260
Study Section
Special Emphasis Panel (ZRG1-GGG-Q (40))
Project Start
Project End
Budget Start
2014-05-10
Budget End
2015-04-30
Support Year
1
Fiscal Year
2014
Total Cost
$301,457
Indirect Cost
$15,479
Name
Brandeis University
Department
Type
DUNS #
616845814
City
Waltham
State
MA
Country
United States
Zip Code
02454
Polleys, Erica J; House, Nealia C M; Freudenreich, Catherine H (2017) Role of recombination and replication fork restart in repeat instability. DNA Repair (Amst) 56:156-165
Lovett, Susan T (2017) Template-switching during replication fork repair in bacteria. DNA Repair (Amst) 56:118-128
Kim, Jane C; Harris, Samantha T; Dinter, Teresa et al. (2017) The role of break-induced replication in large-scale expansions of (CAG)n/(CTG)n repeats. Nat Struct Mol Biol 24:55-60
Neil, Alexander J; Kim, Jane C; Mirkin, Sergei M (2017) Precarious maintenance of simple DNA repeats in eukaryotes. Bioessays 39:
McGinty, Ryan J; Rubinstein, Rachel G; Neil, Alexander J et al. (2017) Nanopore sequencing of complex genomic rearrangements in yeast reveals mechanisms of repeat-mediated double-strand break repair. Genome Res 27:2072-2082
Laranjo, Laura T; Gross, Stephen J; Zeiger, Danna M et al. (2017) SSB recruitment of Exonuclease I aborts template-switching in Escherichia coli. DNA Repair (Amst) 57:12-16
Nguyen, Jennifer H G; Viterbo, David; Anand, Ranjith P et al. (2017) Differential requirement of Srs2 helicase and Rad51 displacement activities in replication of hairpin-forming CAG/CTG repeats. Nucleic Acids Res 45:4519-4531
McGinty, Ryan J; Puleo, Franco; Aksenova, Anna Y et al. (2017) A Defective mRNA Cleavage and Polyadenylation Complex Facilitates Expansions of Transcribed (GAA)n Repeats Associated with Friedreich's Ataxia. Cell Rep 20:2490-2500
Alexander, Jessica L; Beagan, Kelly; Orr-Weaver, Terry L et al. (2016) Multiple mechanisms contribute to double-strand break repair at rereplication forks in Drosophila follicle cells. Proc Natl Acad Sci U S A 113:13809-13814
Beagan, Kelly; McVey, Mitch (2016) Linking DNA polymerase theta structure and function in health and disease. Cell Mol Life Sci 73:603-15

Showing the most recent 10 out of 28 publications