Insulin and glucose regulate the expression of the DNA repair enzyme XPD

Nucleotide excision repair (NER) of damaged DNA is operated by a complex network of DNA repair enzymes that include a protein termed xeroderma pigmentosum complementation group D (XPD). We have previously reported that the expression of XPD is regulated by activation of the insulin receptor and that mutations of the tyrosine kinase domain of the receptor inhibit the insulin-dependent increase in XPD messenger RNA (mRNA) and protein levels. In the present study, we characterize the insulin-dependent signaling pathway leading to the control of XPD expression. Using Chinese hamster ovary (CHO) cells transfected with the human insulin receptor, we demonstrated that the effect of insulin on XPD mRNA levels was mediated via the RAS-signaling and the p70 S6 kinase pathways. On the other hand, the intracellular level of XPD protein was under the exclusive control of the activation of the RAS-dependent cascade in response to insulin. We also studied the effect of acute and chronic exposures to different concentrations of glucose on the insulin-dependent regulation of intracellular XPD levels. A short-term exposure (48 h) to increasing concentrations of glucose potentiated the insulin-dependent regulation of XPD, and this was associated with an efficient protection against glucose-dependent damage to cellular DNA, as determined by the comet assay. Conversely, in cells that were grown for 3 weeks in the presence of glucose concentration greater than 10 mM, the capability of insulin to regulate the level of XPD was significantly reduced, and this promoted a glucose-dependent accumulation of products of DNA damage. In conclusion, glucose and insulin are important regulators of XPD, and prolonged exposure to toxic levels of glucose reduces the insulin-dependent regulation of DNA repair.