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Loss of MUTYH function in human cells leads to accumulation of oxidative damage and genetic instability

Oncogene volume 32, pages 4500–4508 (2013)Cite this article

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Abstract

The DNA glycosylase MUTYH (mutY homolog (Escherichia coli)) counteracts the mutagenic effects of 8-oxo-7,8-dihydroguanine (8-oxodG) by removing adenine (A) misincorporated opposite the oxidized purine. Biallelic germline mutations in MUTYH cause the autosomal recessive MUTYH-associated adenomatous polyposis (MAP). Here we designed new tools to investigate the biochemical defects and biological consequences associated with different MUTYH mutations in human cells. To identify phenotype(s) associated with MUTYH mutations, lymphoblastoid cell lines (LCLs) were derived from seven MAP patients harboring missense as well as truncating mutations in MUTYH. These included homozygous p.Arg245His, p.Gly264TrpfsX7 or compound heterozygous variants (p.Gly396Asp/Arg245Cys, p.Gly396Asp/Tyr179Cys, p.Gly396Asp/Glu410GlyfsX43, p.Gly264TrpfsX7/Ala385ProfsX23 and p.Gly264TrpfsX7/Glu480del). DNA glycosylase assays of MAP LCL extracts confirmed that all these variants were defective in removing A from an 8-oxoG:A DNA substrate, but retained wild-type OGG1 activity. As a consequence of this defect, MAP LCLs accumulated DNA 8-oxodG in their genome and exhibited a fourfold increase in spontaneous mutagenesis at the PIG-A gene compared with LCLs from healthy donors. They were also hypermutable by KBrO3—a source of DNA 8-oxodG—indicating that the relatively modest spontaneous mutator phenotype associated with MUTYH loss can be significantly enhanced by conditions of oxidative stress. These observations identify accumulation of DNA 8-oxodG and a mutator phenotype as likely contributors to the pathogenesis of MUTYH variants.

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Acknowledgements

The authors are grateful to Drs M Fornasarig and D Barana for patient recruitment. This work has been supported by grants to MB from NIH/ISS, AIRC and by Ministero della Salute Alleanza contro il Cancro to AV and MB.

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Author notes

  1. V Ruggieri

    Present address: 6Current address: Laboratory of Pre-Clinical and Translational Research, IRCCS, Referral Cancer Center of Basilicata, Rionero in Vulture (Pz), Italy.,

  2. V Ruggieri and E Pin: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Environment and Primary Prevention, Istituto Superiore di Sanità, Roma, Italy

    V Ruggieri, M T Russo, F Barone, A Minoprio, E Turco, F Mazzei & M Bignami

  2. Division of Experimental Oncology 1, Centro di Riferimento Oncologico, IRCCS, Aviano, Italy

    E Pin, M Quaia & A Viel

  3. Department of Translational Oncology, Istituto Nazionale per la Ricerca sul Cancro (IST-CBA), Genova, Italy

    P Degan

  4. Department of Cell Biology and Neuroscience, Istituto Superiore di Sanità, Rome, Italy

    M Sanchez

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Correspondence to M Bignami.

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Ruggieri, V., Pin, E., Russo, M. et al. Loss of MUTYH function in human cells leads to accumulation of oxidative damage and genetic instability. Oncogene 32, 4500–4508 (2013). https://doi.org/10.1038/onc.2012.479

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