Identification of human MutY homolog (hMYH) as a repair enzyme for 2-hydroxyadenine in DNA and detection of multiple forms of hMYH located in nuclei and mitochondria

Influence of Oxidized Purine Processing on Strand Directionality of Mismatch Repair

“…We show that, in extracts of human cells or Xenopus laevis eggs, the MMR excision machinery can use strand breaks arising upon MYHcatalyzed removal of adenines from G O /A mispairs as initiation sites for exonucleolytic degradation of error-containing strands on circular heteroduplex substrates. A similar phenomenon was not observed either on G O /C-containing heteroduplexes or on substrates containing 2-hydroxyadenine (A O ), another product of purine oxidation (12), even though A O /C and A O /G mispairs were reported previously to be addressed by MYH (23,24).…”

“…MYH has been reported to excise (in addition to adenine from mispairs with G O ) also the oxidation product of adenine, A O , from mispairs with guanine or cytosine (23,24). We therefore included the covalently closed A O /G and A O /C substrates in this assay.…”

MUTYH promotes oxidative microglial activation and inherited retinal degeneration

“…Retinal protein (30 μg) was subjected to SDS-PAGE and transferred onto a polyvinylidene difluoride membrane. The membranes were blocked with Starting Block T20 Blocking Buffer (Thermo Fisher Scientific) for 30 min and then incubated with rabbit anti-hMUTYH antibody, as previously described, (1:3,500) at 4°C overnight (59). After being washed with Tris-buffered saline containing 0.1% Tween, the membranes were incubated with anti-rabbit secondary antibody labeled with horseradish peroxidase (Envision + System, Dako), and signals were developed by chemiluminescence with SuperSignal West Femto (Thermo Fisher Scientific).…”

Oxidative DNA damage in disease—Insights gained from base excision repair glycosylase‐deficient mouse models

“…In human cohorts, mutations in MUTY are associated with colorectal cancer and polyposis, a recessively inherited condition presenting with multiple colorectal adenomas [Al-Tassan et al, 2002;Parker et al, 2002;Chow et al, 2004;Isidro et al, 2004;Dallosso et al, 2008;Casorelli et al, 2010;Kundu et al, 2010;Oka and Nakabeppu, 2011;Mazzei et al, 2013]. The human isoform of MUTY has been demonstrated to have both nuclear and mitochondrial localization, with intracellular localization being associated with specific stages of the cell cycle [Ohtsubo et al, 2000;Boldogh et al, 2001].…”