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hemMascot Server 2.4.0Mascot Parser 2.4.3.0Permanent Cell Line CellCell CultureCell lines: We used two Chinese hamster cell lines (CHO): CHO-DRA10 and CHO-AT3-2 for mutagenesis analysis of Na+/K+ pump ATPase and adenine phosphoribosyltransferase (APRT) loci. Cells were maintained at 37°C with 95% air/5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum. Radioactive labeling of cells: [Methyl-3H]TdR (cat # TRK 637; 45,0 Ci/mmol) purchased from Amersham (GE Healthcare Europe, Orsay, France) was diluted in regular medium. Cells were treated in exponential and asynchronous growth at the same concentration (0.1 mM) but at various specific activities (0.05 to 5 µCi/ml). Cells were incubated for 20 h at 37°C with 5% CO2; at this time, incorporation plateaued, and greater than 95% of cells contained labeled nucleotides. The radioactive medium was subsequently discarded, and cells were washed twice in phosphate-buffered saline (PBS) before processing (1). Experiments were repeated at least thrice. External radiation exposure: Cells were irradiated in exponential and asynchronous growth with a medical 60Co γ-rays external source. Each dose was delivered either on 2 minutes (high rate) or 20 hours (Low rate) in an specific incubator (37°C with 5% CO2). Dosimetry was evaluated inside the incubator. Protein extraction: Cells (10 to 20.106 cells) were washed in PBS, and the cell pellet was re-suspended in lysis buffer (20 mM HEPES, 1 mM DTT, 0.5% NP40, a protease inhibitor cocktail (complete mini, Roche), and a phosphatase inhibitor cocktail (Sigma)). Following 20 min at 4°C, the suspension was centrifuged at 10000 × g for 10 min. The pellet containing cell nuclei was homogenized in solubilization buffer (9 M urea, 4% CHAPS, 0.05% Triton X100, 65 mM DTT and protease inhibitor cocktail). Proteins were then solubilized and quantified as previously described (4). Two-dimensional electrophoresis: Two-dimensional electrophoresis was performed with independent biological experiments and at least three replicates using precast 18-cm strips at pH range 6 to 11 (GE Healthcare) for the first dimension and 12% acrylamide SDS-polyacrylamide gel for the second dimension (5, 6). Analytical gels were performed with 100 µg of protein and stained with Lava-purple (Serva) as recommended by the manufacturer and scanned to images with a Typhoon 9400 (GE). A preparative gel was performed for each condition with 250 µg of protein and stained using a MS compatible silver staining protocol (7). Image analysis Images from analytical gels were analyzed using the Samespots software v4.0 (Non-linear Dynamics, UK). Gel replicates were grouped to create a global analysis with all conditions. Spots of each samples were compared between control and irradiated conditions. A multivariate statistical analysis was performed using the statistic mode of the Samespots software (Non-linear Dynamics, UK). Spots with significant differences (ANOVA t-test p<0.05) were first chosen. Then, only spots with a q-value < 0.05 and a power > 0.8 were finally selected. Spots of interest were selected for subsequent protein identification by mass spectrometry analysis and were picked up using the corresponding preparative silver stained gels. LC MS/MS analysis: Spots were washed with 300 µL of water and then 300 µL of 25 mM NH4HCO3. Destaining was performed twice in the presence of 300 µL of 50% acetonitrile in 25 mM NH4HCO3. Gel pieces were then dehydrated twice by 300 µL of 100% CH3CN and finally dried at 37°C for 1 h. Eight microliters of a trypsin solution (Sequencing Grade Modified Trypsin, Promega, Madison, USA) at a concentration of 0.0125 µg/µL in 25 mM NH4HCO3 was added to every spot. Digestion was performed overnight at 37°C and was stopped by addition of 2 µL of 2 % formic acid. Digests were sonicated in an ultrasonic bath for 10 minutes and supernatants were transferred into HPLC polypropylene tubes. The protein digests were analysed using a Q-TOF mass spectrometer (Maxis ; Bruker Daltonik ), coupled to a nano-chromatography system (HPLC 1200, Agilent Technologies, ) interfaced with an HPLC-Chip system (Chip Cube, Agilent Technologies). Samples were concentrated onto the online pre-column (Agilent Technologies, part G4240-62006, Zorbax 300SB-C18, 40 nL, 5 μm particle size) at a flow rate of 4 µL/min using 0.1% formic acid. After pre-concentration, peptides were separated with a reversed-phase capillary column (Agilent, Zorbax 300SB-C18, 0.075x150mm) at a flow rate of 0.3 µL/min using a 2 steps gradient (3 % to 27 % acetonitrile in 14 min then 27 % to 72 % in 5 min), and eluted directly into the mass spectrometer. The instrument was operated in the positive ion mode with a capillary voltage of 1.8-2.1kV, and a dry gas flow rate of 4 L/min at 140°C. After an initial MS scan at 2 Hz, an IDA mode was employed over the mass range of 300-1400 Th, for the five most intense precursors with an active exclusion of 0.15 min after 2 spectra.PrideBernard S. LopezmaXisUMR CNRS 8200, Institut Gustave Roussy, Université Paris-Saclay, Villejuif, FranceCOMPLETE27406380 Saintigny Y, Chevalier F, Bravard A, Dardillac E, Laurent D, Hem S, Dépagne J, Radicella JP, Lopez BS. A threshold of endogenous stress is required to engage cellular response to protect against mutagenesis. Sci Rep. 2016 Jul 11;6:29412 10.1038/srep29412INRABernard.LOPEZ@gustaveroussy.frMass SpectrometryGel-based experimentMalignant Neoplasm Of OvaryOxidative stressLow dosesMutagenesisProteomicEndogenous stressTritiumhttp://www.ebi.ac.uk/pride/archive/projects/PXD003542Not availableCarbamidomethylOxidationMS/MS raw data were analysed using Compass DataAnalysis software v.4.0.275.0 (Bruker Daltonik GmbH, Bremen, Germany) to generate the peak lists. The Swissprot database (release 2015_03, 547 964 sequences) was queried locally using the Mascot search engine v.2.4.0 (Matrix Science, http://www.matrixscience.com) and with the following parameters : Rodentia for the taxonomy, trypsin as enzyme, 1 missed cleavage allowed, carbamidomethylation of Cystein as fixed modification, oxidation of Methionine as variable modification. Mass tolerance was set to 10 ppm on full scans and 0.05 Da for fragment ions. Proteins were validated once they contained at least two peptide with a p value <0.05. When identified, contaminants proteins (such as keratine) were removed from the identification list and only the two top hit remaining proteins with similar scores and emPAI were considered.ProteomicsCricetulus Griseus (chinese Hamster) (cricetulus Barabensis Griseus)sonia.hem@supagro.inra.frBiologicalFrance10.6019/PXD003542Endogenous stress represents a major source of genome instability, but is in essence difficult to apprehend. Incorporation of labeled radionuclides into DNA constitutes a tractable model to analyze cellular responses to endogenous attacks. Here we show that incorporation of [(3)H]thymidine into CHO cells generates oxidative-induced mutagenesis, but, with a peak at low doses. Proteomic analysis showed that the cellular response differs between low and high levels of endogenous stress. In particular, these results confirmed the involvement of proteins implicated in redox homeostasis and DNA damage signaling pathways. Induced-mutagenesis was abolished by the anti-oxidant N-acetyl cysteine and plateaued, at high doses, upon exposure to L-buthionine sulfoximine, which represses cellular detoxification. The [(3)H]thymidine-induced mutation spectrum revealed mostly base substitutions, exhibiting a signature specific for low doses (GC > CG and AT > CG). Consistently, the enzymatic activity of the base excision repair protein APE-1 is induced at only medium or high doses. Collectively, the data reveal that a threshold of endogenous stress must be reached to trigger cellular detoxification and DNA repair programs; below this threshold, the consequences of endogenous stress escape cellular surveillance, leading to high levels of mutagenesis. Therefore, low doses of endogenous local stress can jeopardize genome integrity more efficiently than higher doses.A threshold of endogenous stress is required to engage cellular response to protect against mutagenesis.Saintigny Yannick Y, Chevalier François F, Bravard Anne A, Dardillac Elodie E, Laurent David D, Hem Sonia S, Dépagne Jordane J, Radicella J Pablo JP, Lopez Bernard S BSsodium salt, Dnahc8, CPD photolyase activity, ammonium formate, 13C-labeled, Size, MeCN, cadmium salt, dAmph, PhrB photolyase activity, APRTD, Particle, FEZL, growth and development, SDH, Basodexan, magnesium formate, 5'-Adenosine monophosphate, Phosphoribosyltransferase, CG18315, 5'-adenylic acid, zinc salt, Long Term, dmTAF[[II]]230, SDS, Polypeptides, protein polypeptide chains, damph, domestic cat, Exposure, 12-trihydroxy-24-oxocholan-24-yl)amino)propyl)-, cobalt(II) formate dihydrate, CH3-C#N, ampicillin acid, AP, B1, Line, Adenosine-5'-monophosphoric acid, Urea, aminobenzylpenicillin, 3, Ampicillin, Software Engineering, NUP96, germacrene A synthase activity, 7, outer pigmented layer of retina, 2210418N07, epithelium, bacterial catalase-peroxidase activity, WMS, ASD2, Nucleus, CG10850, ampicillin anhydrous, Divorced, TFIID TAF250, sarcoma of breast, cel, rya-r44F, CT21282, Chip, ChIP, N, Salt, CHIP, proteins, Divorces, adenosine phosphate, D17Mit170, 5'-Adenylic acid, HPLC, SUPPRESSOR OF AUXIN RESISTANCE 3, Rya-R44F, AI047805, PA1, chip, TRK1, haem catalase activity, pigment epithelium of retina, dehydrated, CG10844, ampicilina, column, D1, n, pigmented retina epithelium, Transphosphoribosidase, TRKA, TrkA, s, Homo sapiens disease, Chinese hamsters, autolysin activity, Nucleotide, phosphatase, 12-trihydroxy-24-oxocholan-24-yl)amino)-, nickel salt, SGS, AI315345, equilase activity, dTAF[[II]]230, Radiation, l(2)k04405, (2S, BZRP, CHO, cobalt (+2) salt, Chinese Hamster Ovary cell, BcDNA:LP07986, TAF200, Longterm Effect, Hst6.7b, GASP, breast sarcoma, Tl3, GAST, Tl2, A330009E03Rik, Software Tools, PBS, Cell Nuclei, ACMICD, PBR, Computer Applications, sodium (4:1:1) salt, Dm1, magnesium salt, cho, CHAPS, Computer Applications Software, Silver, dipyrimidine photolyase (photosensitive), Sizes, 3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, D-(-)-6-(alpha-aminophenylacetamido)penicillanic acid, Tkr, 6-trans-farnesyl-diphosphate diphosphate-lyase (germacrene-A-forming) activity, 5beta, Hydrogen Oxide, ACGNAT, N-dimethyl-N-(3-sulfopropyl)-3-(((3alpha, Software Applications, fast rate, instrument, ATEM1, Source Software, proportionality, formate, Trk-A, rate, cromium (+3), study protocol, TM3, beta Trypsin, Akrylamid, TM5, Apc5, APC5, 7alpha, Applications, Painful bladder syndrome, Taf250, Pyrophosphorylase, hscp30, bacteriolytic toxin activity, ammonium (4:1) salt, spindle cells, methyl cyanide, DROCATHPO, lead (+2) salt, bs36h11.y1, TAF230, Computer Software Applications, BPBS, HSPABP2, DmelCG8604, F10B6_15, Particle Sizes, nickel formate dihydrate, lead formate, Effects, sarcoma of the breast, aluminum salt, number, Nitrate Staining, IMAGE, 2-trans, prop-2-enamide, Carbamide, SCAR16, Silver Nitrate Staining, protein-containing complex, body system, P1-Loop, CG7826, F10B6.15, deoxyribodipyrimidine photolyase activity, stratum pigmentosum retinae, CrAT, period, SPOT, DmelCG18315, Electrophoreses, TITAN 1, At-3, DmelCG5203, manganese catalase activity, TOF, Cricetulus barabensis griseus, CG7835, Gene Products, CG42273, disease or disorder, system, Carmol, spot, C80751, Southern Europe, Application, ZNF312, Open Source Softwares, proportion, anatomical systems, dTAF[[II]]250, NY-CO-7, LATE EMBRYOGENESIS ABUNDANT 1, Longterm, thallium (+1) salt, cell, Software Application, GEA1, Monosodium Salt, beta-Trypsin, rubidium salt, Open Source Software, F23A5.3, 4733401P19Rik, pk18, DmelCG10850, Long-Term, methanoic acid, MBR, Solution, NEM1, Computer Software Application, dTAF250, DmelCG6871, ATPase, Tools, EMB133, PTBR, peptidos, DmelCG10844, 6beta-[(2R)-2-amino-2-phenylacetamido]-2, UBOX1, TM30nm, Chromatographies, 4-(2-hydroxyethyl)-, PTHB1, N-dimethyl-3-sulfo-N-(3-(((3alpha, APRTase, inhibitors, DMCATHPO, DRR, cou, heme catalase activity, 1-(14)C-labeled, DRY, TRK, Trk, Proteins, disorders, CG5203, BG:DS00004.13, potassium formate, copper (+2) salt, backward, acetonitrile, l(2)04405, buffer, (+)-(10R)-germacrene A synthase activity, Cell, CHAMPIGNON, Tool, dTAF230, polypeptide, AMP, Amp, TUBULIN FOLDING COFACTOR D, Lr, native protein, carbonyldiamide, dry, C18, ME-IV., trk, ATIII, Felis domesticus, cupric formate, chemical analysis, Long Term Effects, Mutageneses, CG8604, TAF[[II]]250/230, condition, MFS1, 2-Propenamide, aluminum formate, Adenosine 5'-phosphate, CS1, AMP:pyrophosphate phospho-D-ribosyltransferase, Taf[[II]]250, AU020952, 6R)-6-{[(2R)-2-amino-2-phenylethanoyl]amino}-3, TM-5, lithium formate, RPE, l(2)k00424, holin, hdhc9, ABPC, Ldb, LDB, adenosine-5'P, Separations, pA, postnatal growth, SWDS, 2-dimethylpenam-3alpha-carboxylic acid, CG6871, HEL-189, ethanenitrile, mDRC, retinal pigment epithelium, Longterm Effects, ME-IV, p. pigmentosa retinae, plan specification, Computer Programs, Gene Proteins, ZFP312, l(2)k04913, ARABIDOPSIS THALIANA LATE EMBRYOGENESIS ABUNDANT 1, Applications Softwares, Bra, quotient, deoxyribonucleate pyrimidine dimer lyase (photosensitive), phosphates, quantitative, AT3D, Adenylic acid, reversed, acrylamide, TPMsk3, NCMe, determination, CFTD, postnatal development, instrument configuration, OK|SW-cl.5, zinc formate, Cs-1, HEPES Monosodium Salt, lead salt, protein, PAdo, Pi, lysis, pigmented epithelium, peptide, 6-(D-(2-amino-2-phenylacetamido))-3, hydroxide, Incubator, ammonium tetraformate, Triton, ethylenecarboxamide, peptido, diseases, Min, diseases and disorders, protein aggregate, Effect, Nereid, 2310040B03Rik, Computer Program, DmelCG42273, At3, HEPES Monosodium, SCA3, me75, human disease, MJD, 3-DIMETHYL-7-OXO-4-THIA-1-AZABICYCLO[3.2.0]HEPTANE-2-CARBOXYLIC ACID, reference sample, peptides, TACHD, bA325O24.3, E927b, Open, ATX3, pigmented retina, min, Computer Programs and Programming, mAPC, IBP, OK/SW-cl.5, C6orf207, copper, SCAN, T1, DNA cyclobutane dipyrimidine photolyase activity, phosphate d'adenosine, AT3, lithium salt, 1-Piperazineethanesulfonic acid, SOLO DANCERS, 2.4.2.7, dya, C16orf53, AW046544, necrosis, blood capillary, Bladder pain syndrome, GPHYSD2, Long-Term Effects, ammonium (2:1) salt, inner salt, ratio, catalase-peroxidase activity, Anhydrous ampicillin, (+)-germacrene A synthase activity, PRE, uvrc homolog protein, Ida, IDA, THPH7, Nuclei, deoxyribonucleic cyclobutane dipyrimidine photolyase activity, Dmel_CG7826, AI836084, TAFII-250, 2.3.1.7, pigmented retinal epithelium, TAF250/230, aprt, ur, Source Softwares, AI195249, Programs, Program, carbamide, TAFII250, 3-dimethyl-7-oxo-4-thia-1-azabicyclo(3.2.0)heptane-2-carboxylic acid, catalase reaction, retinal pigment, APRT, C85684, Computer Applications Softwares, fosfato de adenosina, TM30, bacteriocin activity, bA325O24.4, Softwares, Cricetulus aureus, GAS, nickel (+2) salt, GAT, retinal pigment layer, Dmel_CG7835, 5R, SDCCAG7, Lines, Mnb, MNB, ammonium salt, Painful Bladder Syndrome, dLdb, growth pattern, FDR adjusted p-value, RyR, non-developmental growth, retinal pigmented epithelium, DMEM, hydrogen-peroxide:hydrogen-peroxide oxidoreductase activity, TSEPA, cobaltous formate, 6R)-6-{[(2R)-2-AMINO-2-PHENYLETHANOYL]AMINO}-3, MASS, gas, AW124434, 5'-O-phosphonoadenosine, CG17603, Felis silvestris catus, TAF[[II]], ampicillinum, Saint Pierre and Miquelon, EMBRYO DEFECTIVE 133, organ system, disease, Silver Nitrate, H2NC(O)NH2, phr A photolyase activity, lysin activity, DNA-photoreactivating enzyme, Adenine, phosphate, phosphate ions, RYR, Ryr, SR3-5, MJD1, copper salt, St. Pierre and Miquelon, GUANINE NUCLEOTIDE EXCHANGE FACTOR FOR ADP RIBOSYLATION FACTORS 1, Polypeptide, l(3)SH7, HEPES, adenosini phosphas, U00145, time, 1728, Cricetus griseus, Bladder Pain Syndrome, DAMP, Cas1, other disease, d230, Carnitine acetylase, C6orf190, amph, ryr, Bath, cesium salt, 2210017D18Rik, dCHIP, FBN, Gene, dTAFII250, Computer, photoreactivating enzyme activity, adenosine 5'-(dihydrogen phosphate), EfW1, presence, Damp, Buffer, formic acid, Karbamid, Northern Europe, method, potassium salt, polypeptide chain, dmTAF1, Taf230, ECTOL1, 14C-labeled, method used in an experiment, Miquelon and St. Pierre, Harnstoff, RyRs, Chinese hamster, stratum pigmentosa retinae, TFC D, Low, caperase activity, Separated, 6R)-6-{[(2R)-2-amino-2-phenylacetyl]amino}-3, Pro-Mega, TAF250, 5'-AMP, Taf200, MOS3, deoxyribocyclobutadipyrimidine pyrimidine-lyase activity, Adenylate, Staining, PRECOCIOUS, mmol, Acrylamide, inhibiteur, CatA, CATA, Taf1p, labeling, calcium formate, non-neoplastic, OCTD, Miquelon and Saint Pierre, HEL-S-82p, ampicilline, inhibidor, Catl, Suspension, AMP Pyrophosphorylase, Ado5'P, DmelCG3924, disorder, cats, Long-Term Effect, N-2-Hydroxyethylpiperazine-N'-2'-ethanesulfonic Acid, cromium (+3) salt, 12alpha)-3, TAF, CAPM1, 0610033N24Rik, FEZ, p140-TrkA, statistical analysis, sodium formate, F23A5_3, Adenosine 5'-monophosphate, MTC, Controlled, D-MEM, 1-Propanaminium, Applications Software, ADENOSINE MONOPHOSPHATE, 7alpha)-3, Open Source, DYRK1, Controlling, TAF[[II]]250, Computer Software, nickel formate, endonuclease cho, DBI, 3H-labeled, protein complex, cholic acid sulfobetaine1-Propanaminium, 6-trans-farnesyl-diphosphate diphosphate-lyase [(+)-germacrene-A-forming] activity, Cell Lines, strontium formate, l(3)84Ab, inhibitor, EM1 PROTEIN, medical condition, strontium salt, Carnitine acetyltransferase, Peptide, development, ampicillin, high rate, count in organism, stratum pigmentosum (retina), Software Tool, MODIFIER OF SNC1, capillary vessel, PKBS, natural protein, p230, AI114908, Protein, 3-(3-cholamidopropyl)dimethylammoniumpropane sulfonate, JOS, Western Europe, deoxyribonucleic photolyase activity, l(3)63Fb, TFIID, Dyrk1, CAT, connected anatomical system, D-(-)-ampicillin, Software, ACETONITRILE, Corsica, cyanomethane, WMS2, optidase activity, VSD1, Separation, Cas-1, 10^[-9], TAF[[II]]230, dRyR, uree, photolyase activity, urea, CC1, Engineering, Tripcellim, N 2 Hydroxyethylpiperazine N' 2' ethanesulfonic Acid, phosphoric monoester hydrolase activity, TAF[II]250, cat, CGI-97, CG3924, chromic formate, UREA, Protein Gene Products, Trypure, DmelCG17603, dLDB/Chip, Cat01, concentration, SSKS, calcium salt, Peptid, assay, growth, Ultrasonic, TAF1, presence or absence in organism, Rya-r76CDreactivity, Mutageneses., response, responsivityBru, IPP2A2, 2-amino-4-(methylsulfanyl)butanoic acid, Raw, ion, AW549739, cleavage, L Isomer, FBN, Capybara, Gene, rodent, Computer, PHAPII, 5730420M11Rik, peptide, Polypeptides, L-Isomer Methionine, Methionine, Drug Tolerance, ClvPrd, peptido, template-activating factor I, ECTOL1, Gene Products, L-Methionine, Del(8)44H, Software Engineering, Computer Program, Pedameth, WMS, Application, C79691, Svc, Open Source Softwares, Search Engines, SET, peptides, TAF-I, Software Application, M, iones, Open, phosphatase 2A inhibitor I2PP2A, ipp2a2, beta-Trypsin, Open Source Software, Computer Programs and Programming, 2pp2a, Sciences, proteins, Castor Beaver, ions, CG10574, Computer Software Application, Lccp, OCTD, DmelCG4299, mKIAA0989, set, IGAAD, 2-amino-4-(methylthio)butanoic acid, 2PP2A, Tools, 10^[-6], DmelCG10574, taf-ibeta, ppm, Jerboas, dSET, dSet, peptidos, metionina, GPHYSD2, Racemethionine, drug tolerance, SGS, Applications Software, Hydrochaeris, phapii, Open Source, data, Computer Software, DL-Methionine, immune system tolerance, rodents, Proteins, 2-Amino-4-(methylthio)butyric acid, igaad, StF-IT-1, Search, L-Isomer, Source Softwares, Peptide, Engine, Software Tools, Tool, Programs, Dipodidae, ACMICD, polypeptide, Program, Computer Applications, Hydrochaeri, Software Tool, Immune Tolerance, Ionen, I-2PP2A, Computer Applications Software, Dm I-2, Protein, I2PP2A, Computer Applications Softwares, Immunologic Tolerance, Methionin, MFS1, methionine, Softwares, Hmet, Software, Data Base, Met, WMS2, Software Applications, Col4a-1, HLA-DR-associated protein II, DI-2, Protein., Beavers, Source Software, I-2Dm, Engineering, Tripcellim, MASCOT, Rodents, Jerboa, MASS, CG4299, inhibitor of granzyme A-activated DNase, Beaver, beta Trypsin, Rodent, Capybaras, I-2PP1, Protein Gene Products, Computer Programs, Trypure, Gene Proteins, dSET/TAF-Ibeta, Applications, 2610030F17Rik, Applications Softwares, Self Tolerance, TAF-IBETA, Tolerance, Ion, Rodentias, SSKS, alpha-amino-gamma-methylmercaptobutyric acid, Peptid, TAF-Ibeta, Polypeptide, AA407739, Liquimeth, i2pp2a, Computer Software Applications, Immunological Toleranced230, biological signaling, APEN, Cysteine Hydrochloride, DNS, Activity, (Deoxyribonucleotide)n, Gene, dTAFII250, Sulfoximine, protein, protein-containing complex, EfW1, mitochondrial, Deoxyribonucleic acids, 2-amino-4-(S-butylsulfonimidoyl)-, Deoxythymidine, dmTAF[[II]]230, protein polypeptide chains, TITAN 1, Deoxyribonucleic Acid, polypeptide chain, dmTAF1, Taf230, responsivity, 4.2.99.18, Gene Products, DNA Damage Response, TFC D, Low, Half-Cystine, protein aggregate, TAF250, Genomes., reactivity, Taf200, thymus nucleic acid, me75, DNA-(apurinic or apyrimidinic site) lyase, Injury, dTAF[[II]]250, TFIID TAF250, Genomes, cel, cell, L Cysteine, bases, Basen, Double Stranded, Taf1p, Deoxyribonucleic acid, proteins, inhibition of homeostatic process, D17Mit170, T1, dTAF250, signaling process, EMB133, negative regulation of homeostatic process, homeostasis, Double-Stranded DNA, (Deoxyribonucleotide)m, deoxyribonucleic acids, Half Cystine, DNAn, TAF, single organism signaling, Autoregulation, AP endonuclease 1, data, dTAF[[II]]230, TAF[[II]]250, uvrc homolog protein, cou, endonuclease cho, CHO, Zinc Cysteinate, Nucleobase, protein complex, Chinese Hamster Ovary cell, DNAn+1, Proteins, TAF200, incomplete, l(3)84Ab, BG:DS00004.13, Double-Stranded, TAFII-250, TAF250/230, Tl3, Tl2, CHAMPIGNON, Cell, results, dTAF230, Programs, (Deoxyribonucleotide)n+m, abolished, APE-1, TAFII250, TUBULIN FOLDING COFACTOR D, Lr, native protein, positive regulation of homeostatic process, natural protein, cho, p230, Protein, Base2, 2'-Deoxythymidine, proteomic analysis, Base1, Mutageneses, TAF[[II]]250/230, TFIID, ds-DNA, desoxyribose nucleic acid, regulation of homeostatic process, Redox factor-1, 2' Deoxythymidine, Taf[[II]]250, DNA Injury, Genotoxic Stress, DNA Injuries, TAF[[II]]230, Butanoic acid, 3.1.-.-, DNA Lesion, TAF[II]250, L-Cysteine, APEX nuclease, CG17603, TAF[[II]], EMBRYO DEFECTIVE 133, signalling, Protein Gene Products, Gene Proteins, DNA Lesions, DmelCG17603, activation of homeostatic process, Genotoxic, signalling process, Taf250, SR3-5, ds DNA, Stress, Base, Desoxyribonukleinsaeure, Bra, REF-1, nucleobases, DNA, response, Apurinic-apyrimidinic endonuclease 1, General activity, TAF230, Buthionine, TAF1, basebiological signaling, APEN, Cysteine Hydrochloride, DNS, Activity, (Deoxyribonucleotide)n, Gene, Sulfoximine, protein, protein-containing complex, mitochondrial, Deoxyribonucleic acids, 2-amino-4-(S-butylsulfonimidoyl)-, Deoxythymidine, protein polypeptide chains, Deoxyribonucleic Acid, polypeptide chain, responsivity, 4.2.99.18, Gene Products, DNA Damage Response, Low, Half-Cystine, protein aggregate, CHO Cell, Genomes., reactivity, thymus nucleic acid, me75, DNA-(apurinic or apyrimidinic site) lyase, Injury, Genomes, L Cysteine, bases, Basen, Double Stranded, Deoxyribonucleic acid, proteins, inhibition of homeostatic process, D17Mit170, T1, signaling process, negative regulation of homeostatic process, homeostasis, Double-Stranded DNA, (Deoxyribonucleotide)m, deoxyribonucleic acids, Half Cystine, DNAn, single organism signaling, Autoregulation, AP endonuclease 1, data, cou, CHO, Zinc Cysteinate, Nucleobase, protein complex, DNAn+1, Proteins, incomplete, Double-Stranded, Tl3, Tl2, Cell, results, Programs, (Deoxyribonucleotide)n+m, abolished, APE-1, Lr, native protein, positive regulation of homeostatic process, natural protein, Protein, Base2, 2'-Deoxythymidine, proteomic analysis, Base1, Mutageneses, ds-DNA, desoxyribose nucleic acid, regulation of homeostatic process, Redox factor-1, 2' Deoxythymidine, DNA Injury, Genotoxic Stress, DNA Injuries, Butanoic acid, 3.1.-.-, DNA Lesion, L-Cysteine, APEX nuclease, signalling, Protein Gene Products, Gene Proteins, DNA Lesions, activation of homeostatic process, Genotoxic, signalling process, ds DNA, Cells, Stress, Base, Desoxyribonukleinsaeure, Bra, REF-1, nucleobases, DNA, response, Apurinic-apyrimidinic endonuclease 1, General activity, Buthionine, baseMutageneses.383trueProteomic and cellular responses against mutagenesisEndogenous stress represents a major source of genome instability, but is in essence difficult to apprehend. Incorporation of labeled radionuclides into DNA constitutes a tractable model to analyze cellular responses to endogenous attacks. Here we show that incorporation of [3H]thymidine into CHO hamster cells generates oxidative-induced mutagenesis, but, with a peak at low doses. Proteomic analysis showed that the cellular global response differs between low and high levels of endogenous stress. In particular, these results confirmed the involvement of proteins implicated in redox homeostasis and DNA damage signaling pathways. Induced-mutagenesis was abolished by the anti-oxidant N-acetyl cysteine and plateaued, at high doses, upon exposure to L-buthionine sulfoximine, which represses cellular detoxification. The [3H]thymidine-induced mutation spectrum revealed mostly base substitutions, exhibiting a signature specific for low doses (GC>CG and AT>CG). Consistently, the enzymatic activity of the base excision repair protein APE-1 is induced at only medium or high doses. Collectively, the data reveal that a threshold of endogenous stress must be reached to trigger cellular detoxification and DNA repair programs; below this threshold, the consequences of endogenous stress escape cellular surveillance, leading to high levels of mutagenesis. Therefore, low doses of endogenous local stress can jeopardize genome integrity more efficiently than higher 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