Oxidation status of human OGG1-S326C polymorphic variant determines cellular DNA repair capacity.
The hOGG1 gene encodes the DNA glycosylase that removes the mutagenic lesion 7,8-dihyro-8-oxoguanine (8-oxoG) from DNA. A frequently found polymorphism resulting in a serine to cysteine substitution at position 326 of the OGG1 protein has been associated in several molecular epidemiologic studies with cancer development. To investigate whether the variant allele encodes a protein with altered OGG1 function, we compared the 8-oxoG repair activity, both in vivo and in cell extracts, of lymphoblastoid cell lines established from individuals carrying either Ser/Ser or Cys/Cys genotypes. We show that cells homozygous for the Cys variant display increased genetic instability and reduced in vivo 8-oxoG repair rates. Consistently, their extracts have an almost 2-fold lower basal 8-oxoG DNA glycosylase activity when compared with the Ser variant. Treatment with reducing agents of either the Cys variant cells directly or of protein extracts from these cells increases the repair capacity to the level of the Ser variant, whereas it does not affect the activity in cells or extracts from the latter. Furthermore, the DNA glycosylase activity of cells carrying the Cys/Cys alleles is more sensitive to inactivation by oxidizing agents when compared with that of the Ser/Ser cells. Analysis of the redox status of the OGG1 protein in the cells confirms that the lower activity of OGG1-Cys326 is associated with the oxidation of Cys326 to form a disulfide bond. Our findings support the idea that individuals homozygous for the OGG1-Cys variant could more readily accumulate mutations under conditions of oxidative stress.
Bravard
Anne
A
CEA, Institut de Radiobiologie Cellulaire et Moléculaire, UMR217 Centre National de la Recherche Scientifique/CEA, Fontenay aux Roses, France.
Vacher
Monique
M
Moritz
Eva
E
Vaslin
Laurence
L
Hall
Janet
J
Epe
Bernd
B
Radicella
J Pablo
JP
D000483:Alleles; D002460:Cell Line; D045647:DNA Glycosylases / Q000235:genetics* / Q000378:metabolism*; D004260:DNA Repair / Q000502:physiology*; D006801:Humans; D008214:Lymphocytes / Q000201:enzymology / Q000502:physiology; D010084:Oxidation-Reduction; D020641:Polymorphism, Single Nucleotide
D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human