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

V. Ruggieri, E. Pin, M. T. Russo, F. Barone, P. Degan, M. Sanchez, M. Quaia, A. Minoprio, E. Turco, F. Mazzei, A. Viel, M. Bignami

Research output: Contribution to journalArticlepeer-review

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 KBrO 3 - 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.

Original languageEnglish
Pages (from-to)4500-4508
Number of pages9
JournalOncogene
Volume32
Issue number38
DOIs
Publication statusPublished - Sep 19 2013

Keywords

  • 8-oxodG
  • DNA glycosylase activity
  • MUTYH variants
  • MUTYH-associated polyposis
  • PIG-A

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

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