Lipid peroxidation (LPO) products are relatively stable and abundant metabolites, which accumulate in tissues of mammals with aging, being able to modify all cellular nucleophiles, creating protein and DNA adducts including crosslinks. Here, we used cells and mice deficient in the ERCC1-XPF endonuclease required for nucleotide excision repair and the repair of DNA interstrand crosslinks to ask if specifically LPO-induced DNA damage contributes to loss of cell and tissue homeostasis. Ercc1 −/− mouse embryonic fibroblasts were more sensitive than wild-type (WT) cells to the LPO products: 4-hydroxy-2-nonenal (HNE), crotonaldehyde and malondialdehyde. ERCC1-XPF hypomorphic mice were hypersensitive to CCl 4 and a diet rich in polyunsaturated fatty acids, two potent inducers of endogenous LPO. To gain insight into the mechanism of how LPO influences DNA repair-deficient cells, we measured the impact of the major endogenous LPO product, HNE, on WT and Ercc1 −/− cells. HNE inhibited proliferation, stimulated ROS and LPO formation, induced DNA base damage, strand breaks, error-prone translesion DNA synthesis and cellular senescence much more potently in Ercc1 −/− cells than in DNA repair-competent control cells. HNE also deregulated base excision repair and energy production pathways. Our observations that ERCC1-deficient cells and mice are hypersensitive to LPO implicates LPO-induced DNA damage in contributing to cellular demise and tissue degeneration, notably even when the source of LPO is dietary polyunsaturated fats.
|Original language||English (US)|
|Number of pages||18|
|Journal||Free Radical Biology and Medicine|
|State||Published - Aug 20 2018|
Bibliographical noteFunding Information:
This work was supported by the Polish Ministry of Science and Higher Education , grant. N N303 819540 (BT) and by the National Institutes of Health , grants P01-AG043376 (LJN), R01-ES016114 (LJN), P30-AG024837 (LJN), R03-CA121411 (LJN), K99/R00-AG049126 (AUG) and the Ellison Medical Foundation AG-NS-0303-05 (LJN).
Experiments were carried out with the use of CePT infrastructure financed by the European Union – the European Regional Development Fund (Innovative economy 2007–2013, Agreement POIG.02.02.00-14-024/08-00 ). The authors would like to thank Prof. Leon H. F. Mullenders for providing Xpa −/− cells and Aleksander Chlebowski for his help in microscopy studies.
This work was supported by the Polish Ministry of Science and Higher Education, grant. N N303 819540 (BT) and by the National Institutes of Health, grants P01-AG043376 (LJN), R01-ES016114 (LJN), P30-AG024837 (LJN), R03-CA121411 (LJN), K99/R00-AG049126 (AUG) and the Ellison Medical Foundation AG-NS-0303-05 (LJN).
- DNA damage
- Lipid peroxidation