TY - JOUR

T1 - Response of PRIMPOL-Knockout Human Lung Adenocarcinoma A549 Cells to Genotoxic Stress

AU - Gromova, Anastasia S

AU - Boldinova, Elizaveta O

AU - Kim, Daria V

AU - Chuprov-Netochin, Roman N

AU - Leonov, Sergey V

AU - Pustovalova, Margarita V

AU - Zharkov, Dmitry O

AU - Makarova, Alena V

N1 - This work was supported by the Russian Science Foundation (grant no. 18-14-00354 for A.V.M.; generation of PRIMPOL cells, analysis of metabolic activity and DNA-replicating cell fractions, cell cycle analysis) and the Russian Foundation for Basic Research (grant no. 20-34-90092-Aspirants for D.V.K.; generation of clonal cell lines), and by the Ministry of Science and Higher Education of the Russian Federation (State Assignment 075-03-2023-106, project no. FSMG-2023-0015 for M.V.P.; cell treatment with ionizing radiation).

PY - 2023/11

Y1 - 2023/11

N2 - Human DNA primase/polymerase PrimPol synthesizes DNA primers de novo after replication fork stalling at the sites of DNA damage, thus contributing to the DNA damage tolerance. The role of PrimPol in response to the different types of DNA damage is poorly understood. We knocked out the PRIMPOL gene in the lung carcinoma A549 cell line and characterized the response of the obtained cells to the DNA damage caused by hydrogen peroxide, methyl methanesulfonate (MMS), cisplatin, bleomycin, and ionizing radiation. The PRIMPOL knockout reduced the number of proliferating cells and cells in the G2 phase after treatment with MMS and caused a more pronounced delay of the S phase in the cisplatin-treated cells. Ionizing radiation at a dose of 10 Gy significantly increased the content of apoptotic cells among the PRIMPOL-deficient cells, while the proportion of cells undergoing necroptosis increased in both parental and knockout cells at any radiation dose. The viability of PRIMPOL-deficient cells upon the hydrogen peroxide-induced oxidative stress increased compared to the control cells, as determined by the methyl tetrazolium (MTT) assay. The obtained data indicate the involvement of PRIMPOL in the modulation of adaptive cell response to various types of genotoxic stress.

AB - Human DNA primase/polymerase PrimPol synthesizes DNA primers de novo after replication fork stalling at the sites of DNA damage, thus contributing to the DNA damage tolerance. The role of PrimPol in response to the different types of DNA damage is poorly understood. We knocked out the PRIMPOL gene in the lung carcinoma A549 cell line and characterized the response of the obtained cells to the DNA damage caused by hydrogen peroxide, methyl methanesulfonate (MMS), cisplatin, bleomycin, and ionizing radiation. The PRIMPOL knockout reduced the number of proliferating cells and cells in the G2 phase after treatment with MMS and caused a more pronounced delay of the S phase in the cisplatin-treated cells. Ionizing radiation at a dose of 10 Gy significantly increased the content of apoptotic cells among the PRIMPOL-deficient cells, while the proportion of cells undergoing necroptosis increased in both parental and knockout cells at any radiation dose. The viability of PRIMPOL-deficient cells upon the hydrogen peroxide-induced oxidative stress increased compared to the control cells, as determined by the methyl tetrazolium (MTT) assay. The obtained data indicate the involvement of PRIMPOL in the modulation of adaptive cell response to various types of genotoxic stress.

KW - Humans

KW - DNA-Directed DNA Polymerase/metabolism

KW - A549 Cells

KW - Cisplatin/pharmacology

KW - Hydrogen Peroxide/pharmacology

KW - DNA Replication

KW - DNA Damage

KW - Adenocarcinoma of Lung/genetics

KW - DNA Primase/genetics

KW - Multifunctional Enzymes/genetics

KW - replication

KW - DNA damage

KW - apoptosis

KW - PRIMPOL

KW - primase

KW - knockout cell lines

KW - damage tolerance

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85178198231&origin=inward&txGid=21f6f7c89b919f7bd096642a44fc873d

UR - https://www.mendeley.com/catalogue/970ee5b9-caad-3c43-b2d6-cb370e8d0c86/

U2 - 10.1134/S0006297923110214

DO - 10.1134/S0006297923110214

M3 - Article

C2 - 38105210

VL - 88

SP - 1933

EP - 1943

JO - Biochemistry (Moscow)

JF - Biochemistry (Moscow)

SN - 0006-2979

IS - 11

ER -