The goals of Project 2 are to develop a detailed picture of human disease-associated RecQ helicase protein function, and to use this information to delineate the role of these RecQ helicases in human tumor cell proliferation, survival and the response to chemotherapy. In research to date we have focused on functional analyses of the human Werner syndrome protein WRN. In the proposed research we extend these analyses to encompass all three disease-associated human RecQ helicases, the WRN, BLM and RECQL4 proteins associated with the heritable deficiency/genetic instability/cancer predisposition syndromes Werner syndrome (WRN), Bloom syndrome (BLM) and Rothmund-Thomson (RTS) syndrome, respectively.
Aim 1. Develop a comprehensive proteomic description of the human disease-associated WRN, BlM and RECQl4 human RecQ helicases. We will identify functionally important RecQ helicase protein associations and post-translational modifications (PTM's) by using quantitative affinity purification mass spectrometry, shRNA-mediated epistatic interaction mapping and functional analyses of post-translational modification (PTM) site mutants.
Aim 2. Determine functional requirement for WRN, BlM and RECQL4 in DNA metabolism. We will determine the role of each of these RecQ helicases in DNA replication, translesion DNA synthesis (TLS), homology-dependent recombination (HDR) and DNA break repair using high resolution assays.
Aim 3. Determine functional redundancy among WRN, BlM and RECQL4 in DNA metabolism, and clinically relevant synthetic interactions with additional genes/proteins, drugs or small molecules. We will identify unique and redundant functions of the 'disease-associated human RecQ helicases, and synthetic interactions between disease-associated human RecQ helicases, their interacting proteins, and chemotherapeutic agents, drugs or small molecules that can beused to selectively kill RecQ-deficient human tumors.

Public Health Relevance

The human RecQ helicase proteins maintain genomic stability and protect against DNA damage and DNA metabolic errors. The proposed research will provide insight into the functions of these critical proteins in human cells, and how RecQ function is related to cancer risk and the response to cancer chemotherapy.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA077852-15
Application #
8494583
Study Section
Special Emphasis Panel (ZCA1-GRB-S)
Project Start
Project End
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
15
Fiscal Year
2013
Total Cost
$253,892
Indirect Cost
$71,027
Name
University of Washington
Department
Type
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Reid-Bayliss, Kate S; Loeb, Lawrence A (2017) Accurate RNA consensus sequencing for high-fidelity detection of transcriptional mutagenesis-induced epimutations. Proc Natl Acad Sci U S A 114:9415-9420
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Poole, William; Leinonen, Kalle; Shmulevich, Ilya et al. (2017) Multiscale mutation clustering algorithm identifies pan-cancer mutational clusters associated with pathway-level changes in gene expression. PLoS Comput Biol 13:e1005347
Fu, Wenqing; Ligabue, Alessio; Rogers, Kai J et al. (2017) Human RECQ Helicase Pathogenic Variants, Population Variation and ""Missing"" Diseases. Hum Mutat 38:193-203
Beckman, Robert A; Loeb, Lawrence A (2017) Evolutionary dynamics and significance of multiple subclonal mutations in cancer. DNA Repair (Amst) 56:7-15
Fox, Edward J; Salk, Jesse J; Loeb, Lawrence A (2016) Exploring the implications of distinct mutational signatures and mutation rates in aging and cancer. Genome Med 8:30
Tokita, Mari; Kennedy, Scott R; Risques, Rosa Ana et al. (2016) Werner syndrome through the lens of tissue and tumour genomics. Sci Rep 6:32038
Reid-Bayliss, Kate S; Arron, Sarah T; Loeb, Lawrence A et al. (2016) Why Cockayne syndrome patients do not get cancer despite their DNA repair deficiency. Proc Natl Acad Sci U S A 113:10151-6

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