WRN Exonuclease activity is blocked by specific oxidatively induced base lesions positioned in either DNA strand

Werner syndrome (WS) is a premature aging disorder caused by mutations in the WS gene (WRN). Although WRN has been suggested to play an important role in DNA metabolic pathways, such as recombination, replication and repair, its precise role still remains to be determined. WRN possesses ATPase, helicase and exonuclease activities. Previous studies have shown that the WRN exonuclease is inhibited in vitro by certain lesions induced by oxidative stress and positioned in the digested strand of the substrate. The presence of the 70/86 Ku heterodimer (Ku), participating in the repair of double-strand breaks (DSBs), alleviates WRN exonuclease blockage imposed by the oxidatively induced DNA lesions. The current study demonstrates that WRN exonuclease is inhibited by several additional oxidized bases, and that Ku stimulates the WRN exonuclease to bypass these lesions. Specific lesions present in the non-digested strand were shown also to inhibit the progression of the WRN exonuclease; however, Ku was not able to stimulate WRN exonuclease to bypass these lesions. Thus, this study considerably broadens the spectrum of lesions which block WRN exonuclease progression, shows a blocking effect of lesions in the non-digested strand, and supports a function for WRN and Ku in a DNA damage processing pathway

Oxidation products of 5-methyl cytosine are decreased in senescent cells and tissues of progeroid mice

5-Hydroxymethylcytosine and 5-formylcytosine are stable DNA base modifications generated from 5-methylcytosine by the ten-eleven translocation protein family that function as epigenetic markers. 5-Hydroxymethyluracil may also be generated from thymine by ten-eleven translocation enzymes. Here, we asked if these epigenetic changes accumulate in senescent cells, since they are thought to be inversely correlated with proliferation. Testing this in ERCC1-XPF-deficient cells and mice also enabled discovery if these DNA base changes are repaired by nucleotide excision repair. Epigenetic marks were measured in proliferating, quiescent and senescent wild-type (WT) and Ercc1−/− primary mouse embryonic fibroblasts. The pattern of epigenetic marks depended more on the proliferation status of the cells than their DNA repair capacity. The cytosine modifications were all decreased in senescent cells compared to quiescent or proliferating cells, whereas 5-(hydroxymethyl)-2′-deoxyuridine was increased. In vivo, both 5-(hydroxymethyl)-2′-deoxyuridine and 5-(hydroxymethyl)-2′-deoxycytidine were significantly increased in liver tissues of aged WT mice compared to young adult WT mice. Livers of Ercc1-deficient mice with premature senescence and aging had reduced level of 5-(hydroxymethyl)- 2′-deoxycytidine and 5-formyl-2′-deoxycytidine compared to aged-matched WT controls. Taken together, we demonstrate for the first time, that 5-(hydroxymethyl)-2′-deoxycytidine is significantly reduced in senescent cells and tissue, potentially yielding a novel marker of senescence

Spontaneous DNA damage to the nuclear genome promotes senescence, T redox imbalance and aging

Accumulation of senescent cells over time contributes to aging and age-related diseases. However, what drives senescence in vivo is not clear. Here we used a genetic approach to determine if spontaneous nuclear DNA damage is sufficient to initiate senescence in mammals. Ercc1-/Δ mice with reduced expression of ERCC1-XPF endonuclease have impaired capacity to repair the nuclear genome. Ercc1-/Δ mice accumulated spontaneous, oxidative DNA damage more rapidly than wild-type (WT) mice. As a consequence, senescent cells accumulated more rapidly in Ercc1-/Δ mice compared to repair-competent animals. However, the levels of DNA damage and senescent cells in Ercc1-/Δ mice never exceeded that observed in old WT mice. Surprisingly, levels of reactive oxygen species (ROS) were increased in tissues of Ercc1-/Δ mice to an extent identical to naturally-aged WT mice. Increased enzymatic production of ROS and decreased antioxidants contributed to the elevation in oxidative stress in both Ercc1-/Δ and aged WT mice. Chronic treatment of Ercc1-/Δ mice with the mitochondrial-targeted radical scavenger XJB-5–131 attenuated oxidative DNA damage, senescence and age-related pathology. Our findings indicate that nuclear genotoxic stress arises, at least in part, due to mitochondrial-derived ROS, and this spontaneous DNA damage is sufficient to drive increased levels of ROS, cellular senescence, and the consequent age-related physiological decline

Different faces of DNA repair - Nobel price 2015 in chemistry

W 2015 r. Nagroda Nobla w dziedzinie chemii została przyznana za badania mechanistyczne nad naprawą DNA Paulowi Modrichowi, Tomasowi Lindahlowi i Azizowi Sancarowi. Paul Modrich pracuje w Howard Hughes Medical Institute oraz Duke University School of Medicine, Durham, USA. Nagrodą zostały wyróżnione jego prace nad naprawą źle dopasowanych zasad, które powstają głównie podczas replikacji, zaś ten typ naprawy jest "pierwszą linią ochrony" stabilności genomu. Tomas Lindahl jest profesorem chemii medycznej i fizycznej, emerytowanym dyrektorem Cancer Research UK London Research Institute, Clare Hall Laboratories, South Mimms, Wielka Brytania. Nagrodę Nobla otrzymał za odkrycia w dziedzinie naprawy przez wycięcie zasady usuwającej z DNA niewielkie uszkodzenia, głównie oksydacyjne i alkilacyjne. Aziz Sancar jest profesorem biochemii i biofizyki na University of North Carolina School of Medicine, Chapel Hill, USA. Nagrodę Nobla otrzymał za osiągnięcia w dziedzinie naprawy przez wycięcie nukleotydu. System ten usuwa z DNA duże modyfikacje takie jak dimery pirymidynowe indukowane światłem ultrafioletowym. Badania uczonych stworzyły podwaliny pod zrozumienie mechanizmu ewolucji świata ożywionego, a także procesów nowotworowych i opracowanie nowoczesnych terapii.The Nobel Prize in chemistry for 2015 was awarded to Paul Modrich, Tomas Lindahl and Aziz Sancar for mechanistic studies on DNA repair. Paul Modrich works in Howard Hughes Medical Institute and Duke University School of Medicine, Durham, USA. The prize has been awarded for his work on Mismatch Repair, which removes mismatched nucleotides formed mainly during replication and is the "first line of defense" of genome stability. Tomas Lindahl is a professor of medical and physical chemistry, emeritus director of Cancer Research UK London Research Institute, Clare Hall Laboratories, South Mimms, United Kingdom. The Nobel Prize was awarded to him for discoveries on Base Excision Repair, which removes from the DNA small lesions, mainly alkylated and oxidatively formed damages. Aziz Sancar is a professor in biochemistry and biophysics at University of North Carolina School of Medicine, Chapel Hill, USA. He was awarded for the achievements on Nucleotide Excision Repair. The system removes from the DNA big lesions, such as pyrimidine dimers induced by ultraviolet light. Studies of these researchers made a basis for understanding of the evolution of living world as well as carcinogenic process and for elaboration of novel therapies

Damage of DNA and proteins by major lipid peroxidation products in genome stability

Oxidative stress and lipid peroxidation (LPO) accompanying infections and chronic inflammation may induce several human cancers. LPO products are characterized by carbohydrate chains of different length, reactive aldehyde groups and double bonds, which make these molecules reactive to nucleic acids, proteins and cellular thiols. LPO-derived adducts to DNA bases form etheno-type and propano-type exocyclic rings, which have profound mutagenic potential, and are elevated in several cancer-prone diseases. Adducts of long chain LPO products to DNA bases inhibits transcription. Elimination from DNA of LPO-induced lesions is executed by several repair systems: base excision repair (BER), direct reversal by AlkB family proteins, nucleotide excision repair (NER) and recombination. Modifications of proteins with LPO products may regulate cellular processes like apoptosis, cell signaling and senescence. This review summarizes consequences of LPO products presence in cell, particularly 4-hydroxy-2-nonenal in terms of genomic stability

Oxidatively damaged DNA and its repair in colon carcinogenesis

Inflammation, high fat, high red meat and low fiber consumption have for long been known as the most important etiological factors of sporadic colorectal cancers (CRC). Colon cancer originates from neoplastic transformation in a single layer of epithelial cells occupying colonic crypts, in which migration and apoptosis program becomes disrupted. This results in the formation of polyps and metastatic cancers. Mutational program in sporadic cancers involves APC gene, in which mutations occur most abundantly in the early phase of the process. This is followed by mutations in RAS, TP53, and other genes. Progression of carcinogenic process in the colon is accompanied by augmentation of the oxidative stress, which manifests in the increased level of oxidatively damaged DNA both in the colon epithelium, and in blood leukocytes and urine, already at the earliest stages of disease development. Defence mechanisms are deregulated in CRC patients: (i) antioxidative vitamins level in blood plasma declines with the development of disease; (ii) mRNA level of base excision repair enzymes in blood leukocytes of CRC patients is significantly increased; however, excision rate is regulated separately, being increased for 8-oxoGua, while decreased for lipid peroxidation derived ethenoadducts, Ade and Cyt; (iii) excision rate of Ade and Cyt in colon tumors is significantly increased in comparison to asymptomatic colon margin, and ethenoadducts level is decreased. This review highlights mechanisms underlying such deregulation, which is the driving force to colon carcinogenesis