A Multi-Endpoint Approach to Base Excision Repair Incision Activity Augmented by PARylation and DNA Damage Levels in Mice: Impact of Sex and Age

• Published in: International journal of molecular sciences 2020-09, Vol.21 (18), p.6600
• Main Authors: Winkelbeiner, Nicola, Wandt, Viktoria K, Ebert, Franziska, Lossow, Kristina, Bankoglu, Ezgi E, Martin, Maximilian, Mangerich, Aswin, Stopper, Helga, Bornhorst, Julia, Kipp, Anna P, Schwerdtle, Tanja
• Format: Article
• Language: English
• Subjects: Adenosine diphosphate; ADP-ribosylation; Age; ageing; Aging; Aging - metabolism; Animals; Base excision repair; base excision repair (incision activity); Denaturing Gradient Gel Electrophoresis; Deoxyguanosine; Deoxyribonucleic acid; DNA; DNA Damage; DNA damage response; DNA Repair; Enzymes; Female; Hep G2 Cells; Homeostasis; Humans; Impact damage; Liver; Liver - metabolism; maintenance of genomic integrity; Male; Mammals; Mice, Inbred C57BL; Oligonucleotides - isolation & purification; Physiology; Poly ADP Ribosylation; Proteins; Repair; sex; Sex Characteristics
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21186600

Investigation of processes that contribute to the maintenance of genomic stability is one crucial factor in the attempt to understand mechanisms that facilitate ageing. The DNA damage response (DDR) and DNA repair mechanisms are crucial to safeguard the integrity of DNA and to prevent accumulation of persistent DNA damage. Among them, base excision repair (BER) plays a decisive role. BER is the major repair pathway for small oxidative base modifications and apurinic/apyrimidinic (AP) sites. We established a highly sensitive non-radioactive assay to measure BER incision activity in murine liver samples. Incision activity can be assessed towards the three DNA lesions 8-oxo-2'-deoxyguanosine (8-oxodG), 5-hydroxy-2'-deoxyuracil (5-OHdU), and an AP site analogue. We applied the established assay to murine livers of adult and old mice of both sexes. Furthermore, poly(ADP-ribosyl)ation (PARylation) was assessed, which is an important determinant in DDR and BER. Additionally, DNA damage levels were measured to examine the overall damage levels. No impact of ageing on the investigated endpoints in liver tissue were found. However, animal sex seems to be a significant impact factor, as evident by sex-dependent alterations in all endpoints investigated. Moreover, our results revealed interrelationships between the investigated endpoints indicative for the synergetic mode of action of the cellular DNA integrity maintaining machinery.