Germ-line variant of human NTH1 DNA glycosylase induces genomic instability and cellular transformation.
Base excision repair (BER) removes at least 20,000 DNA lesions per human cell per day and is critical for the maintenance of genomic stability. We hypothesize that aberrant BER, resulting from mutations in BER genes, can lead to genomic instability and cancer. The first step in BER is catalyzed by DNA N-glycosylases. One of these, n(th) endonuclease III-like (NTH1), removes oxidized pyrimidines from DNA, including thymine glycol. The rs3087468 single nucleotide polymorphism of the NTH1 gene is a G-to-T base substitution that results in the NTH1 D239Y variant protein that occurs in ∼6.2% of the global population and is found in Europeans, Asians, and sub-Saharan Africans. In this study, we functionally characterize the effect of the D239Y variant expressed in immortal but nontransformed human and mouse mammary epithelial cells. We demonstrate that expression of the D239Y variant in cells also expressing wild-type NTH1 leads to genomic instability and cellular transformation as assessed by anchorage-independent growth, focus formation, invasion, and chromosomal aberrations. We also show that cells expressing the D239Y variant are sensitive to ionizing radiation and hydrogen peroxide and accumulate double strand breaks after treatment with these agents. The DNA damage response is also activated in D239Y-expressing cells. In combination, our data suggest that individuals possessing the D239Y variant are at risk for genomic instability and cancer.
Department of Microbiology and Molecular Genetics, The Markey Center for Molecular Genetics, University of Vermont, Burlington, VT 05405-0068, USA.
Heather A
HA
Galick
Scott
S
Kathe
Minmin
M
Liu
Susan
S
Robey-Bond
Dawit
D
Kidane
Susan S
SS
Wallace
Joann B
JB
Sweasy
D002471:Cell Transformation, Neoplastic / Q000235:genetics; D043224:Deoxyribonuclease (Pyrimidine Dimer) / Q000235:genetics*; D042822:Genomic Instability; D006801:Humans; D020641:Polymorphism, Single Nucleotide
D043224:Deoxyribonuclease (Pyrimidine Dimer); C468347:NTHL1 protein, human
DNA repair; MCF10A breast epithelial cells; mutagenesis