Single Molecule Glycosylase Studies with Engineered 8-oxoguanine DNA Damage Sites Show Functional Defects of a MUTYH Polyposis Variant

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2019 Jan 31

PMID 30698731

Citations 8

Authors

Shane R Nelson

Scott D Kathe

Thomas S Hilzinger

April M Averill

David M Warshaw

Susan S Wallace

Andrea J Lee

Affiliations

Soon will be listed here.

Abstract

Proper repair of oxidatively damaged DNA bases is essential to maintain genome stability. 8-Oxoguanine (7,8-dihydro-8-oxoguanine, 8-oxoG) is a dangerous DNA lesion because it can mispair with adenine (A) during replication resulting in guanine to thymine transversion mutations. MUTYH DNA glycosylase is responsible for recognizing and removing the adenine from 8-oxoG:adenine (8-oxoG:A) sites. Biallelic mutations in the MUTYH gene predispose individuals to MUTYH-associated polyposis (MAP), and the most commonly observed mutation in some MAP populations is Y165C. Tyr165 is a 'wedge' residue that intercalates into the DNA duplex in the lesion bound state. Here, we utilize single molecule fluorescence microscopy to visualize the real-time search behavior of Escherichia coli and Mus musculus MUTYH WT and wedge variant orthologs on DNA tightropes that contain 8-oxoG:A, 8-oxoG:cytosine, or apurinic product analog sites. We observe that MUTYH WT is able to efficiently find 8-oxoG:A damage and form highly stable bound complexes. In contrast, MUTYH Y150C shows decreased binding lifetimes on undamaged DNA and fails to form a stable lesion recognition complex at damage sites. These findings suggest that MUTYH does not rely upon the wedge residue for damage site recognition, but this residue stabilizes the lesion recognition complex.

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References

Mitra S, Boldogh I, Izumi T, Hazra T . Complexities of the DNA base excision repair pathway for repair of oxidative DNA damage. Environ Mol Mutagen. 2001; 38(2-3):180-90. PMC: 4927302. DOI: 10.1002/em.1070. View

Nemec A, Donigan K, Murphy D, Jaeger J, Sweasy J . Colon cancer-associated DNA polymerase β variant induces genomic instability and cellular transformation. J Biol Chem. 2012; 287(28):23840-9. PMC: 3390657. DOI: 10.1074/jbc.M112.362111. View

Wooden S, Bassett H, Wood T, McCullough A . Identification of critical residues required for the mutation avoidance function of human MutY (hMYH) and implications in colorectal cancer. Cancer Lett. 2004; 205(1):89-95. DOI: 10.1016/j.canlet.2003.10.006. View

Friedman J, Stivers J . Detection of damaged DNA bases by DNA glycosylase enzymes. Biochemistry. 2010; 49(24):4957-67. PMC: 2886154. DOI: 10.1021/bi100593a. View

Parker A, Sieber O, Shi C, Hua L, Takao M, Tomlinson I . Cells with pathogenic biallelic mutations in the human MUTYH gene are defective in DNA damage binding and repair. Carcinogenesis. 2005; 26(11):2010-8. DOI: 10.1093/carcin/bgi166. View

Markkanen E, Dorn J, Hubscher U . MUTYH DNA glycosylase: the rationale for removing undamaged bases from the DNA. Front Genet. 2013; 4:18. PMC: 3584444. DOI: 10.3389/fgene.2013.00018. View

Wallace S, Murphy D, Sweasy J . Base excision repair and cancer. Cancer Lett. 2012; 327(1-2):73-89. PMC: 3361536. DOI: 10.1016/j.canlet.2011.12.038. View

Yamtich J, Nemec A, Keh A, Sweasy J . A germline polymorphism of DNA polymerase beta induces genomic instability and cellular transformation. PLoS Genet. 2012; 8(11):e1003052. PMC: 3493456. DOI: 10.1371/journal.pgen.1003052. View

Wallace S . Biological consequences of free radical-damaged DNA bases. Free Radic Biol Med. 2002; 33(1):1-14. DOI: 10.1016/s0891-5849(02)00827-4. View

10.

Kow Y . Mechanism of action of Escherichia coli exonuclease III. Biochemistry. 1989; 28(8):3280-7. DOI: 10.1021/bi00434a024. View

11.

Qi Y, Nam K, Spong M, Banerjee A, Sung R, Zhang M . Strandwise translocation of a DNA glycosylase on undamaged DNA. Proc Natl Acad Sci U S A. 2012; 109(4):1086-91. PMC: 3268267. DOI: 10.1073/pnas.1111237108. View

12.

Hayashi H, Tominaga Y, Hirano S, McKenna A, Nakabeppu Y, Matsumoto Y . Replication-associated repair of adenine:8-oxoguanine mispairs by MYH. Curr Biol. 2002; 12(4):335-9. DOI: 10.1016/s0960-9822(02)00686-3. View

13.

Molatore S, Russo M, DAgostino V, Barone F, Matsumoto Y, Albertini A . MUTYH mutations associated with familial adenomatous polyposis: functional characterization by a mammalian cell-based assay. Hum Mutat. 2009; 31(2):159-66. DOI: 10.1002/humu.21158. View

14.

Colussi C, Parlanti E, Degan P, Aquilina G, Barnes D, MacPherson P . The mammalian mismatch repair pathway removes DNA 8-oxodGMP incorporated from the oxidized dNTP pool. Curr Biol. 2002; 12(11):912-8. DOI: 10.1016/s0960-9822(02)00863-1. View

15.

Liu H, Lu A . Escherichia coli MutY protein has both N-glycosylase and apurinic/apyrimidinic endonuclease activities on A.C and A.G mispairs. Proc Natl Acad Sci U S A. 1992; 89(18):8779-83. PMC: 50004. DOI: 10.1073/pnas.89.18.8779. View

16.

Porello S, Leyes A, David S . Single-turnover and pre-steady-state kinetics of the reaction of the adenine glycosylase MutY with mismatch-containing DNA substrates. Biochemistry. 1998; 37(42):14756-64. DOI: 10.1021/bi981594+. View

17.

Chmiel N, Golinelli M, Francis A, David S . Efficient recognition of substrates and substrate analogs by the adenine glycosylase MutY requires the C-terminal domain. Nucleic Acids Res. 2001; 29(2):553-64. PMC: 29658. DOI: 10.1093/nar/29.2.553. View

18.

Blainey P, Luo G, Kou S, Mangel W, Verdine G, Bagchi B . Nonspecifically bound proteins spin while diffusing along DNA. Nat Struct Mol Biol. 2009; 16(12):1224-9. PMC: 2889498. DOI: 10.1038/nsmb.1716. View

19.

Lee S, Verdine G . Atomic substitution reveals the structural basis for substrate adenine recognition and removal by adenine DNA glycosylase. Proc Natl Acad Sci U S A. 2009; 106(44):18497-502. PMC: 2773975. DOI: 10.1073/pnas.0902908106. View

20.

Sieber O, Lipton L, Crabtree M, Heinimann K, Fidalgo P, Phillips R . Multiple colorectal adenomas, classic adenomatous polyposis, and germ-line mutations in MYH. N Engl J Med. 2003; 348(9):791-9. DOI: 10.1056/NEJMoa025283. View