Depletion of Deoxyribonucleotide Pools Is an Endogenous Source of DNA Damage in Cells Undergoing Oncogene-Induced Senescence

• Published in: The American journal of pathology Vol. 182; no. 1; pp. 142 - 151
• Main Authors: Mannava, Sudha, Moparthy, Kalyana C, Wheeler, Linda J, Natarajan, Venkatesh, Zucker, Shoshanna N, Fink, Emily E, Im, Michael, Flanagan, Sheryl, Burhans, William C, Zeitouni, Nathalie C, Shewach, Donna S, Mathews, Christopher K, Nikiforov, Mikhail A
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 2013; American Society for Investigative Pathology
• Subjects: Cell Proliferation; Cells, Cultured; Cellular Senescence - genetics; Cellular Senescence - physiology; Deoxyribonucleotides - genetics; Deoxyribonucleotides - metabolism; DNA Damage - genetics; DNA Replication - genetics; Fibroblasts - metabolism; Fibroblasts - physiology; Humans; Oncogenes - physiology; Pathology; Proto-Oncogene Proteins p21(ras) - physiology; Regular; Ribonucleotide Reductases - biosynthesis; Ribonucleotide Reductases - physiology; Thymidylate Synthase - biosynthesis; Thymidylate Synthase - physiology
• ISSN: 0002-9440; 1525-2191
• DOI: 10.1016/j.ajpath.2012.09.011

In normal human cells, oncogene-induced senescence (OIS) depends on induction of DNA damage response. Oxidative stress and hyperreplication of genomic DNA have been proposed as major causes of DNA damage in OIS cells. Here, we report that down-regulation of deoxyribonucleoside pools is another endogenous source of DNA damage in normal human fibroblasts (NHFs) undergoing HRASG12V -induced senescence. NHF-HRASG12V cells underexpressed thymidylate synthase (TS) and ribonucleotide reductase (RR), two enzymes required for the entire de novo deoxyribonucleotide biosynthesis, and possessed low dNTP levels. Chromatin at the promoters of the genes encoding TS and RR was enriched with retinoblastoma tumor suppressor protein and histone H3 tri-methylated at lysine 9. Importantly, ectopic coexpression of TS and RR or addition of deoxyribonucleosides substantially suppressed DNA damage, senescence-associated phenotypes, and proliferation arrest in two types of NHF-expressing HRASG12V . Reciprocally, short hairpin RNA-mediated suppression of TS and RR caused DNA damage and senescence in NHFs, although less efficiently than HRASG12V . However, overexpression of TS and RR in quiescent NHFs did not overcome proliferation arrest, suggesting that unlike quiescence, OIS requires depletion of dNTP pools and activated DNA replication. Our data identify a previously unknown role of deoxyribonucleotides in regulation of OIS.