Cancer-associated variants and a common polymorphism of MUTYH exhibit reduced repair of oxidative DNA damage using a GFP-based assay in mammalian cells
Open accessAbstract
Biallelic germline mutations in the base excision repair enzyme gene MUTYH lead to multiple colorectal adenomas and carcinomas referred to as MUTYH-associated polyposis. MUTYH removes adenine misincorporated opposite the DNA oxidation product, 8-oxoguanine (OG), thereby preventing accumulation of G:C to T:A transversion mutations. The most common cancer-associated MUTYH variant proteins when expressed in bacteria exhibit reduced OG:A mismatch affinity and adenine removal activity. However, direct evaluation of OG:A mismatch repair efficiency in mammalian cells has not been assessed due to the lack of an appropriate assay. To address this, we developed a novel fluorescence-based assay of OG:A repair and measured the repair capacity of MUTYH-associated polyposis variants expressed in Mutyh-/- mouse embryonic fibroblasts (MEFs). The repair of a single site-specific synthetic lesion in a green fluorescent protein reporter leads to green fluorescent protein expression with co-expression of a red fluorescent protein serving as the transfection control. Cell lines that stably express the MUTYH-associated polyposis variants G382D and Y165C have significantly lower OG:A repair versus wild-type MEFs and MEFs expressing human wild-type MUTYH. The MUTYH allele that encodes the Q324H variant is found at a frequency above 40% in samples from different ethnic groups and has long been considered phenotypically silent but has recently been associated with increased cancer risk in several clinical studies. In vitro analysis of Q324H MUTYH expressed in insect cells showed that it has reduced enzyme activity similar to that of the known cancer variant G382D. Moreover, we find that OG:A repair in MEFs expressing Q324H was significantly lower than wild-type controls, establishing that Q324H is functionally impaired and providing further evidence that this common variant may lead to increased cancer risk.
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Defects in DNA repair and DNA damage response pathways can lead to human diseases such as cancer and neurodegenerative diseases.
“Defective DNA repair and DDR pathways may lead to several human diseases, such as cancer and neurodegenerative diseases [6, 33–38]”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [34]
Defects in DNA repair and DDR pathways are associated with cancer and neurodegenerative diseases.
“Dysfunctions in these pathways are linked to cancer and neurodegenerative diseases [6, 33–38]”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [34]
Cites
dbSNP: the NCBI database of genetic variation
Sherry, S. T. (2001) · Nucleic Acids Research · doi:10.1093/nar/29.1.308 · Cited by 7,502 · see citing works
Tsien, R. Y. (1998) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.67.1.509 · Cited by 6,178 · see citing works
A comprehensive catalogue of somatic mutations from a human cancer genome
Pleasance, E. D., Cheetham, R. K., Stephens, P. J., McBride, D. J., Humphray, S. J., Greenman, C. D., Varela, I., Lin, M.-L., Ordóñez, G. R., Bignell, G. R., et al. (2010) · Nature · doi:10.1038/nature08658 · Cited by 1,696 · see citing works
Inherited variants of MYH associated with somatic G:C→T:A mutations in colorectal tumors
Tassan, N. A. A., Chmiel, N. H., Maynard, J. H., Fleming, N. I., Livingston, A. L., Williams, G. T., Hodges, A., Davies, D. R., David, S. S., Sampson, J. R., et al. (2002) · Nature Genetics · doi:10.1038/ng828 · Cited by 1,392 · see citing works
Self-citationBase-excision repair of oxidative DNA damage
David, S. S., O'Shea, V. L., & Kundu, S. (2007) · Nature · doi:10.1038/nature05978 · Cited by 1,238 · see citing works
Self-citationA genome-wide association study of global gene expression
Dixon, A. L., Liang, L., Moffatt, M. F., Chen, W., Heath, S. C., Wong, K., Taylor, J. C., Burnett, E. C., Gut, I. G., Farrall, M., et al. (2007) · Nature Genetics · doi:10.1038/ng2109 · Cited by 946 · see citing works
Multiple Colorectal Adenomas, Classic Adenomatous Polyposis, and Germ-Line Mutations inMYH
Sieber, O. M., Lipton, L. R., Crabtree, M., Heinimann, K., Fidalgo, P. O., Phillips, R. K. S., Bisgaard, M.-L., Orntoft, T. F., Aaltonen, L. A., Hodgson, S. V., et al. (2003) · New England Journal of Medicine · doi:10.1056/nejmoa025283 · Cited by 859 · see citing works
The mutY gene: a mutator locus in Escherichia coli that generates G.C----T.A transversions.
Nghiem, Y., Cabrera, M., Cupples, C. G., & Miller, J. H. (1988) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.85.8.2709 · Cited by 343 · see citing works
Structural basis for removal of adenine mispaired with 8-oxoguanine by MutY adenine DNA glycosylase
Fromme, J. C., Banerjee, A., Huang, S. J., & Verdine, G. L. (2004) · Nature · doi:10.1038/nature02306 · Cited by 331 · see citing works
Guan, Y., Manuel, R. C., Arvai, A. S., Parikh, S. S., Mol, C. D., Miller, J. H., Lloyd, R. S., & Tainer, J. A. (1998) · Nature Structural Biology · doi:10.1038/4168 · Cited by 325 · see citing works
Xie, Y., Yang, H., Cunanan, C., Okamoto, K., Shibata, D., Pan, J., Barnes, D. E., Lindahl, T., McIlhatton, M., Fishel, R., et al. (2004) · Cancer Research · doi:10.1158/0008-5472.can-03-3834 · Cited by 291 · see citing works
Self-citationAssociation Between Biallelic and Monoallelic Germline MYH Gene Mutations and Colorectal Cancer Risk
Croitoru, M. E., Cleary, S. P., Nicola, N. D., Manno, M., Selander, T., Aronson, M. D., Redston, M. S., Cotterchio, M., Knight, J. A., Gryfe, R., et al. (2004) · JNCI Journal of the National Cancer Institute · doi:10.1093/jnci/djh288 · Cited by 258 · see citing works
Carcinogenesis in MYH-associated polyposis follows a distinct genetic pathway.
Lipton, L. R., Halford, S. E. R., Johnson, V., Novelli, M. R., Jones, A., Cummings, C., Barclay, E., Sieber, O. M., Sadat, A. T. E., Bisgaard, M.-L., et al. (2003) · PubMed · Cited by 229 · see citing works
The Mammalian Mismatch Repair Pathway Removes DNA 8-oxodGMP Incorporated from the Oxidized dNTP Pool
Colussi, C., Parlanti, E., Degan, P., Aquilina, G., Barnes, D., Macpherson, P., Karran, P., Crescenzi, M., Eugenia Dogliotti, Y., & Bignami, M. (2002) · Current Biology · doi:10.1016/s0960-9822(02)00863-1 · Cited by 228 · see citing works
Parker, A. R., Gu, Y., Mahoney, W., Lee, S., Singh, K. K., & Lu, A.-L. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m008463200 · Cited by 220 · see citing works
Gu, Y., Parker, A. R., Wilson, T., Bai, H., Chang, D., & Lu, A.-L. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m108618200 · Cited by 206 · see citing works
Analysis of MUTYH Genotypes and Colorectal Phenotypes in Patients With MUTYH-Associated Polyposis
Nielsen, M., van de Beld, M. C. J. , Jones, N., Vogt, S. L., Tops, C. M. J., Vasen, H. F. A., Sampson, J. R., Aretz, S., & Hes, F. J. (2008) · Gastroenterology · doi:10.1053/j.gastro.2008.10.056 · Cited by 170 · see citing works
MUTYH-associated polyposis—From defect in base excision repair to clinical genetic testing
Cheadle, J. P., & Sampson, J. R. (2006) · DNA repair · doi:10.1016/j.dnarep.2006.11.001 · Cited by 146 · see citing works
Τheodoratou, E., Campbell, H., Tenesa, A., Houlston, R. S., Webb, E. L., Lubbe, S. J., Broderick, P., Gallinger, S., Croitoru, E. M., Jenkins, M. A., et al. (2010) · British Journal of Cancer · doi:10.1038/sj.bjc.6605966 · Cited by 125 · see citing works
An 8-oxo-guanine repair pathway coordinated by MUTYH glycosylase and DNA polymerase λ
van Loon, B., & Hübscher, U. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0907280106 · Cited by 117 · see citing works
Fleischmann, C., Peto, J., Cheadle, J. P., Shah, B., Sampson, J. R., & Houlston, R. S. (2004) · International Journal of Cancer · doi:10.1002/ijc.20020 · Cited by 114 · see citing works
Cancer risks for monoallelic MUTYH mutation carriers with a family history of colorectal cancer
Win, A. K., Cleary, S. P., Dowty, J. G., Baron, J. A., Young, J. P., Buchanan, D. D., Southey, M. C., Burnett, T. S., Parfrey, P. S., Green, R. C., et al. (2010) · International Journal of Cancer · doi:10.1002/ijc.25870 · Cited by 108 · see citing works
Characterization of Mutant MUTYH Proteins Associated With Familial Colorectal Cancer
Ali, M., Kim, H., Cleary, S. P., Cupples, C. G., Gallinger, S., & Bristow, R. G. (2008) · Gastroenterology · doi:10.1053/j.gastro.2008.04.035 · Cited by 97 · see citing works
Replication-Associated Repair of Adenine:8-Oxoguanine Mispairs by MYH
Hayashi, H., Tominaga, Y., Hirano, S., McKenna, A. E., Nakabeppu, Y., & Matsumoto, Y. (2002) · Current Biology · doi:10.1016/s0960-9822(02)00686-3 · Cited by 94 · see citing works
Kasahara, M., Osawa, K., Yoshida, K., et al. (2008) · Journal of Experimental & Clinical Cancer Research · doi:10.1186/1756-9966-27-49 · Cited by 93 · see citing works
Chmiel, N. H., Livingston, A. L., & David, S. S. (2003) · Journal of Molecular Biology · doi:10.1016/s0022-2836(03)00124-4 · Cited by 92 · see citing works
Self-citationShi, G., Chang, D., Cheng, C.-C., Guan, X., Venclovas, Č., & Lu, A.-L. (2006) · Biochemical Journal · doi:10.1042/bj20060774 · Cited by 91 · see citing works
Shinmura, K. (2000) · Nucleic Acids Research · doi:10.1093/nar/28.24.4912 · Cited by 88 · see citing works
Leiden open variation database of the MUTYH gene
Out, A. A., Tops, C. M. J., Nielsen, M., Weiss, M. M., van Minderhout, I. J. H. M., Fokkema, I. F. A. C., Buisine, M., Claes, K., Colas, C., Fodde, R., et al. (2010) · Human Mutation · doi:10.1002/humu.21343 · Cited by 76 · see citing works
Genomic predictors of interindividual differences in response to DNA damaging agentsPreprint
Fry, R. C., Svensson, J. P., Valiathan, C., Wang, E. Y., Hogan, B. J., Bhattacharya, S., Bugni, J. M., Whittaker, C. A., & Samson, L. D. (2008) · Genes & Development · doi:10.1101/gad.1688508 · Cited by 75 · see citing works
Luncsford, P. J., Chang, D., Shi, G., Bernstein, J., Madabushi, A., Patterson, D. N., Lu, A.-L., & Toth, E. A. (2010) · Journal of Molecular Biology · doi:10.1016/j.jmb.2010.08.045 · Cited by 68 · see citing works
Repair and mutagenic potency of 8-oxoG:A and 8-oxoG:C base pairs in mammalian cells
Page, F. L. (1998) · Nucleic Acids Research · doi:10.1093/nar/26.5.1276 · Cited by 67 · see citing works
Miyaishi, A., Osawa, K., Osawa, Y., et al. (2009) · Journal of Experimental & Clinical Cancer Research · doi:10.1186/1756-9966-28-10 · Cited by 64 · see citing works
Evidence for accelerated colorectal adenoma–carcinoma progression in MUTYH -associated polyposis?
Nieuwenhuis, M. H., Vogt, S. L., Jones, N., Nielsen, M., Hes, F. J., Sampson, J. R., Aretz, S., & Vasen, H. F. A. (2011) · Gut · doi:10.1136/gut.2010.229104 · Cited by 64 · see citing works
Dallosso, A. R., Dolwani, S., Jones, N., Jones, S., Colley, J. P., Maynard, J. H., Idziaszczyk, S., Humphreys, V., Arnold, J., Donaldson, A., et al. (2008) · Gut · doi:10.1136/gut.2007.145748 · Cited by 62 · see citing works
Common variants in human CRC genes as low-risk alleles
Picelli, S., Zając, P., Zhou, X., Edler, D., Lenander, C., Dalén, J., Hjern, F., Lundqvist, N., Lindforss, U., Påhlman, L., et al. (2010) · European Journal of Cancer · doi:10.1016/j.ejca.2010.01.013 · Cited by 61 · see citing works
Pope, M. A., Chmiel, N. H., & David, S. S. (2004) · DNA repair · doi:10.1016/j.dnarep.2004.10.003 · Cited by 56 · see citing works
Self-citationKundu, S., Brinkmeyer, M. K., Livingston, A. L., & David, S. S. (2009) · DNA repair · doi:10.1016/j.dnarep.2009.09.009 · Cited by 53 · see citing works
Self-citationRusso, M. T., Luca, G. D., Casorelli, I., Degan, P., Molatore, S., Barone, F., Mazzei, F., Pannellini, T., Musiani, P., & Bignami, M. (2009) · Cancer Research · doi:10.1158/0008-5472.can-08-3292 · Cited by 52 · see citing works
Xie, Y., Yang, H., Miller, J. H., Shih, D. M., Hicks, G. G., Xie, J., & Shiu, R. P. C. (2008) · Carcinogenesis · doi:10.1093/carcin/bgn033 · Cited by 49 · see citing works
Self-citationShinmura, K., Goto, M., Suzuki, M., et al. (2011) · The Journal of Pathology · doi:10.1002/path.2953 · Cited by 48 · see citing works
Molatore, S., Russo, M. T., D’Agostino, V., Barone, F., Matsumoto, Y., Albertini, A. M., Minoprio, A., Degan, P., Mazzei, F., Bignami, M., et al. (2009) · Human Mutation · doi:10.1002/humu.21158 · Cited by 44 · see citing works
Functional analysis of MUTYH mutated proteins associated with familial adenomatous polyposis
D’Agostino, V., Minoprio, A., Torreri, P., Marinoni, I., Bossa, C., Petrucci, T. C., Albertini, A. M., Ranzani, G. N., Bignami, M., & Mazzei, F. (2010) · DNA repair · doi:10.1016/j.dnarep.2010.03.008 · Cited by 43 · see citing works
Goto, M., Shinmura, K., Nakabeppu, Y., Tao, H., Yamada, H., Tsuneyoshi, T., & Sugimura, H. (2010) · Human Mutation · doi:10.1002/humu.21363 · Cited by 43 · see citing works
Tao, H., Shinmura, K., Suzuki, M., et al. (2008) · Cancer Science · doi:10.1111/j.1349-7006.2007.00694.x · Cited by 42 · see citing works
Livingston, A. L., Kundu, S., Pozzi, M. H., Anderson, D. W., & David, S. S. (2005) · Biochemistry · doi:10.1021/bi050976u · Cited by 42 · see citing works
Self-citationDNA polymerase β and PARP activities in base excision repair in living cells
Masaoka, A., Horton, J. K., Beard, W. A., & Wilson, S. H. (2009) · DNA repair · doi:10.1016/j.dnarep.2009.08.004 · Cited by 40 · see citing works
Rieder, M. J., Livingston, R. J., Stanaway, I. B., & Nickerson, D. A. (2008) · Drug Metabolism Reviews · doi:10.1080/03602530801952880 · Cited by 32 · see citing works
Kundu, S., Brinkmeyer, M. K., Eigenheer, R. A., & David, S. S. (2010) · DNA repair · doi:10.1016/j.dnarep.2010.07.002 · Cited by 27 · see citing works
Self-citationInherited defects in the DNA glycosylase MYH cause multiple colorectal adenoma and carcinoma
Cheadle, J. P., Dolwani, S., & Sampson, J. R. (2003) · Carcinogenesis · doi:10.1093/carcin/bgg068 · Cited by 21 · see citing works
Hahm, S.-H., Park, J.-H., Ko, S. I., Lee, Y.-R., Chung, I.-S., Chung, J.-H., Kang, L., & Han, Y.-S. (2011) · BMB Reports · doi:10.5483/bmbrep.2011.44.5.352 · Cited by 20 · see citing works
Burger, K., Matt, K., Kieser, N., Gebhard, D., & Bergemann, J. (2010) · BMC Biotechnology · doi:10.1186/1472-6750-10-46 · Cited by 16 · see citing works
MUTYH hotspot mutations in unselected colonoscopy patients
Casper, M., Plotz, G., Juengling, B., et al. (2012) · Colorectal Disease · doi:10.1111/j.1463-1318.2012.02920.x · Cited by 13 · see citing works
MUTYHhotspot mutations in unselected colonoscopy patients
Casper, M., Plotz, G., Juengling, B., et al. (2011) · Colorectal Disease · doi:10.1111/j.1463-1318.2011.02920.x · Cited by 4 · see citing works
The 2nd Biennial Scientific Meeting of International Society for Gastrointestinal Hereditary Tumours
Iwama∥, T. (2007) · Familial Cancer · doi:10.1007/s10689-007-9142-z · Cited by 3 · see citing works
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- Authors
- Raetz, A. G., Xie, Y., Kundu, S., Brinkmeyer, M. K., Chang, C., & David, S. S.
- Affiliations
- University of California, Davis (US)
- Health Sciences Centre (CA)
- Corresponding author
- David, Sheila S.
- Venue
- Carcinogenesis
- Publisher
- Oxford University Press (OUP)
- Published
- 2012-08-19
- Volume
- 33
- Issue
- 11
- Pages
- 2301-2309
- ISSN
- 0143-3334 (ISSN-L)
- 1460-2180
- PMC
- PMC3483017
- OpenAlex
- W2165733686
- Cited by
- 45
- References
- 55
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 3 · 2023: 2 · 2022: 2 · 2021: 3 · 2020: 4 · 2019: 1 · 2018: 5 · 2017: 3 · 2016: 3 · 2015: 3 · 2014: 6 · 2013: 4
- FWCI
- 2.99
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Carcinogenesis (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Cancer Institute (CA-67985, R01CA067985)
- National Institute of Environmental Health Sciences (T32 ES007058-33, T32ES007059)
- Topics
- Health Sciences › Medicine › Pathology and Forensic Medicine › Genetic factors in colorectal cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
- Keywords
- MUTYH, MUTYH-associated polyposis, 8-oxoguanine, oxidative DNA damage, base excision repair, DNA mismatch repair, G:C to T:A transversions, colorectal cancer risk, germline mutations
- MeSH terms
- Animals, Biological Assay, Cells, Cultured, DNA Damage, DNA Repair, Embryo, Mammalian, Fibroblasts, Flow Cytometry, Humans, Lung Neoplasms, Mutation, N-Glycosyl Hydrolases, Polymorphism, Genetic, RNA, Messenger, Blotting, Western, Colorectal Neoplasms, Apoptosis, Mice, Knockout, Reverse Transcriptase Polymerase Chain Reaction, DNA Glycosylases, Cell Proliferation, Green Fluorescent Proteins, Mice, Real-Time Polymerase Chain Reaction
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