The MUTYH glycosylase has garnered the spotlight with the discovery of a correlation between inherited MUTYH variations and colorectal cancer (CRC). Using pre-steady kinetics and noncleavable 2'- deoxyadenosine analogue-containing oligonucleotides, we provided information on the functional defects of two missense variants of MUTYH found in colorectal cancer that was key for establishing the connection between MUTYH variants and colorectal cancer (now referred to a MUTYH-associated polyposis or MAP). MUTYH prevents mutations associated with 8-oxo-7,8-dihydro-2'-deoxyguanosine (OG) by removal of misincorporated adenine residues from OG:A mismatches, and thus aberrant functioning MUTYH leads to the accumulation of mutations in tumor suppressor genes, like APC. Since the original discovery of MAP, over 100 different variants of MUTYH have been discovered. Our laboratory has focused on revealing fundamental features of MUTYH and MUTYH variants. Our underlying hypothesis is that a detailed molecular understanding of OG:A mismatch recognition goes hand-in-hand with revealing intricacies of the relationship between MUTYH and colorectal cancer. We have recently developed mammalian cell assays that we will use to evaluate the repair of modified substrates in cells. This will allow us to gauge how catalytic and binding defects impact overall repair in cells. We have recent results with the bacterial enzymes that suggests that the 2-amino- group of OG is critical for initial localization f OG, and we will further test this hypothesis by making additional derivatives and directly monitoring the search process using single-molecule techniques. This will provide important information into features that are involved in the initial damage detection step, and allow for better prediction of features that made compromise damage detection. We also will use our enzymatic assays in conjunction with cellular assays to provide insight into the role of the interdomain connector (IDC) region, and the Zn(II) coordination site we recently discovered on mediating efficient repair, and coordinating with downstream repair proteins, and proteins involved in damage signaling. Lastly, we also will elaborate on work that implicated MUTYH activity in increasing the sensitivity of cells to alkylation damage. As part of this work, we will develop novel methods to tag sites of MutY glycosylase activity in cells. The significance of this work is that it will provide important information as to the how MUTYH dysfunction, either inherited or acquired, may set the stage for carcinogenesis. In addition, the role of MUTYH in mediating cellular responses to oxidative damage and alkylation may provide new strategies to target cancer cells.

Public Health Relevance

This project will provide fundamental information on the functional properties of inherited variations in MUTYH associated with a familial colorectal syndrome, MUTYH-associated polyposis of MAP. This will be useful in evaluating the extent with which alterations predispose individuals to cancer. In addition, this work will also reveal fundamental molecular features of MUTYH-function that if disrupted by oxidative stress may also lead to cancer. In addition, we will test a novel hypothesis that high levels of MUTYH may correlate with a strong response to alkylating chemotherapeutics. Thus, this work will provide important basic information on MUTYH that also translates directly to a clinical setting.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA067985-22
Application #
9388948
Study Section
Synthetic and Biological Chemistry A Study Section (SBCA)
Program Officer
Okano, Paul
Project Start
1995-07-01
Project End
2020-11-30
Budget Start
2017-12-01
Budget End
2018-11-30
Support Year
22
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of California Davis
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
047120084
City
Davis
State
CA
Country
United States
Zip Code
95618
Ha, Yang; Arnold, Anna R; Nuñez, Nicole N et al. (2017) Sulfur K-Edge XAS Studies of the Effect of DNA Binding on the [Fe4S4] Site in EndoIII and MutY. J Am Chem Soc 139:11434-11442
Banda, Douglas M; Nuñez, Nicole N; Burnside, Michael A et al. (2017) Repair of 8-oxoG:A mismatches by the MUTYH glycosylase: Mechanism, metals and medicine. Free Radic Biol Med 107:202-215
Bartels, Phillip L; Zhou, Andy; Arnold, Anna R et al. (2017) Electrochemistry of the [4Fe4S] Cluster in Base Excision Repair Proteins: Tuning the Redox Potential with DNA. Langmuir 33:2523-2530
Manlove, Amelia H; McKibbin, Paige L; Doyle, Emily L et al. (2017) Structure-Activity Relationships Reveal Key Features of 8-Oxoguanine: A Mismatch Detection by the MutY Glycosylase. ACS Chem Biol 12:2335-2344
Wickramaratne, Susith; Banda, Douglas M; Ji, Shaofei et al. (2016) Base Excision Repair of N(6)-Deoxyadenosine Adducts of 1,3-Butadiene. Biochemistry 55:6070-6081
Shen, Yan; McMackin, Marissa Z; Shan, Yuxi et al. (2016) Frataxin Deficiency Promotes Excess Microglial DNA Damage and Inflammation that Is Rescued by PJ34. PLoS One 11:e0151026
Woods, Ryan D; O'Shea, Valerie L; Chu, Aurea et al. (2016) Structure and stereochemistry of the base excision repair glycosylase MutY reveal a mechanism similar to retaining glycosidases. Nucleic Acids Res 44:801-10
Mullins, Elwood A; Shi, Rongxin; Parsons, Zachary D et al. (2015) The DNA glycosylase AlkD uses a non-base-flipping mechanism to excise bulky lesions. Nature 527:254-8
Brinkmeyer, Megan K; David, Sheila S (2015) Distinct functional consequences of MUTYH variants associated with colorectal cancer: Damaged DNA affinity, glycosylase activity and interaction with PCNA and Hus1. DNA Repair (Amst) 34:39-51
Engstrom, Lisa M; Brinkmeyer, Megan K; Ha, Yang et al. (2014) A zinc linchpin motif in the MUTYH glycosylase interdomain connector is required for efficient repair of DNA damage. J Am Chem Soc 136:7829-32

Showing the most recent 10 out of 25 publications