TY - JOUR

T1 - Use of qPCR to Evaluate Efficiency of the Bulky DNA Damage Removal in Extracts of Mammalian Cells with Different Maximum Lifespan

AU - Popov, Aleksei A.

AU - Shamanin, Vladimir A.

AU - Petruseva, Irina O.

AU - Evdokimov, Aleksei N.

AU - Lavrik, Olga I.

N1 - This work was financially supported by the Russian Science Foundation (project no. 19-74-10056-P).

PY - 2024/7

Y1 - 2024/7

N2 - Proteins of nucleotide excision repair system (NER) are responsible for detecting and removing a wide range of bulky DNA damages, thereby contributing significantly to the genome stability maintenance within mammalian cells. Evaluation of NER functional status in the cells is important for identifying pathological changes in the body and assessing effectiveness of chemotherapy. The following method, described herein, has been developed for better assessment of bulky DNA damages removal in vitro, based on qPCR. Using the developed method, NER activity was compared for the extracts of the cells from two mammals with different lifespans: a long-lived naked mole-rat (Heterocephalus glaber) and a short-lived mouse (Mus musculus). Proteins of the H. glaber cell extract have been shown to be 1.5 times more effective at removing bulky damage from the model DNA substrate than the proteins of the M. musculus cell extract. These results are consistent with the experimental data previously obtained. The presented method could be applied not only in fundamental studies of DNA repair in mammalian cells, but also in clinical practice.

AB - Proteins of nucleotide excision repair system (NER) are responsible for detecting and removing a wide range of bulky DNA damages, thereby contributing significantly to the genome stability maintenance within mammalian cells. Evaluation of NER functional status in the cells is important for identifying pathological changes in the body and assessing effectiveness of chemotherapy. The following method, described herein, has been developed for better assessment of bulky DNA damages removal in vitro, based on qPCR. Using the developed method, NER activity was compared for the extracts of the cells from two mammals with different lifespans: a long-lived naked mole-rat (Heterocephalus glaber) and a short-lived mouse (Mus musculus). Proteins of the H. glaber cell extract have been shown to be 1.5 times more effective at removing bulky damage from the model DNA substrate than the proteins of the M. musculus cell extract. These results are consistent with the experimental data previously obtained. The presented method could be applied not only in fundamental studies of DNA repair in mammalian cells, but also in clinical practice.

KW - DNA repair

KW - PCR

KW - longevity

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85201276714&origin=inward&txGid=7a6aa821d0cc4d137924811ff222e5ea

UR - https://www.mendeley.com/catalogue/439332a4-e8aa-32a8-bb44-6dd3117be37b/

U2 - 10.1134/S0006297924070022

DO - 10.1134/S0006297924070022

M3 - Article

C2 - 39218017

VL - 89

SP - 1183

EP - 1191

JO - Biochemistry (Moscow)

JF - Biochemistry (Moscow)

SN - 0006-2979

IS - 7

ER -