Investigating the effects of the presence of foreign DNA on DNA methylation and DNA repair events in cultured eukaryotic cells.
Methylation of DNA in eukaryotic cells, global as well as gene-specific, is affected by endogenous and endogenous factors. In this paper, it is reported that deviations in DNA methylation and expression of genes involved in DNA repair and the cell cycle are affected in 143B cultured cells containing an expression vector. Global DNA methylation analysis with cytosine-extension assay revealed a decreased global DNA methylation in the presence of the expression vector. Less promoter-specific methylation, as measured by bisulfite-MS PCR, was observed for MGMT and p16INK4a in vector-containing cells. Comet assay investigations revealed a negative effect on the DNA repair capacity of both BER and NER in Complex III compromised cells. This was reflected in the down-regulation of hOGG1 and ERCC1 expression. The results presented in this paper support the existence of a strong relationship between impaired mitochondrial function and deviations in DNA methylation and extend this relationship to impaired DNA repair.
Centre of Human Metabonomics, North-West University, Potchefstroom 2520, South Africa. jean.dutoit@nwu.ac.za
J
J
Du Toit
F H
FH
van der Westhuizen
P J
PJ
Pretorius
D002460:Cell Line; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000235:genetics; D004260:DNA Repair; D044127:Epigenesis, Genetic; D005057:Eukaryotic Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D011401:Promoter Regions, Genetic
D019941:Cyclin-Dependent Kinase Inhibitor p16; D015254:DNA Modification Methylases