-
Mutation Research Nov 1976Methyl methanesulfonate (MMS) mutagenesis of Chlamydomonas reinhardtii at different stages of the synchronous cell-cycle revealed the following results. (1) Induction of...
Methyl methanesulfonate (MMS) mutagenesis of Chlamydomonas reinhardtii at different stages of the synchronous cell-cycle revealed the following results. (1) Induction of phenotypically distinct Mendelian (nuclear), str-50 and non-Mendelian (chloroplast) str-500P, streptomycin resistant mutants was relatively high during the first portion of the cell-cycle when chloroplast DNA replication is known to occur. (2) A second and more pronounced interval of enhanced Mendelian, str-50 mutant induction was observed near the middle of the cell-cycle when the initial stages of nuclear DNA replication occur. Induction of non-Mendelian, str-500P mutants was inconsistent during this period. (3) The incidence of mutants from a second phenotypically distinct class of non-Mendelian streptomycin-resistant mutants (str-500D) was not increased over control levels at any stage of the cell-cycle examined. It is concluded that MMS, like N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), may not be the most suitable general mutagen for this alga because its enhanced mutagenesis of cells in the nuclear S phase could result in multiple closely linked mutations.
Topics: Cell Division; Chlamydomonas; DNA Replication; Drug Resistance, Microbial; Mesylates; Methyl Methanesulfonate; Mutation; Streptomycin
PubMed: 187932
DOI: 10.1016/0027-5107(76)90036-1 -
Fungal Biology May 2019DNA damage can cause mutations that in fungal plant pathogens lead to hypervirulence and resistance to pesticides. Almost nothing is known about the response of these...
DNA damage can cause mutations that in fungal plant pathogens lead to hypervirulence and resistance to pesticides. Almost nothing is known about the response of these fungi to DNA damage. We performed transcriptomic and phosphoproteomic analyses of Fusarium oxysporum exposed to methyl methanesulfonate (MMS). At the RNA level we observe massive induction of DNA repair pathways including the global genome nucleotide excision. Cul3, Cul4, several Ubiquitin-like ligases and components of the proteasome are significantly induced. In agreement, we observed drug synergism between a proteasome inhibitor and MMS. While our data suggest that Yap1 and Xbp1 networks are similarly activated in response to damage in yeast and F. oxysporum we were able to observe modules that were MMS-responsive in F. oxysporum and not in yeast. These include transcription/splicing modules that are upregulated and respiration that is down-regulated. In agreement, MMS treated cells are much more sensitive to a respiration inhibitor. At the phosphoproteomic level, Adenylate cyclase, which generates cAMP, is phosphorylated in response to MMS and forms a network of phosphorylated proteins that include cell cycle regulators and several MAPKs. Our analysis provides a starting point in understanding how genomic changes in response to DNA damage occur in Fusarium species.
Topics: DNA Damage; DNA Repair; Fusarium; Gene Expression Profiling; Methyl Methanesulfonate; Mutagens; Phosphoproteins; Plant Diseases; Proteome; Stress, Physiological
PubMed: 31053330
DOI: 10.1016/j.funbio.2019.03.007 -
Chemical Research in Toxicology Oct 2019N7-Methyl-2'-deoxyguanosine (MdG) is the major damage product in DNA produced by methylating agents, but it often thought to be nontoxic and nonmutagenic. MdG is...
N7-Methyl-2'-deoxyguanosine (MdG) is the major damage product in DNA produced by methylating agents, but it often thought to be nontoxic and nonmutagenic. MdG is chemically unstable. An abasic site (AP) is the major product produced from MdG under physiologically relevant conditions. AP formation is frequently considered to be responsible for the cytotoxic effects of MdG, but the reaction is suppressed in nucleosome core particles (NCPs). Recently, it was discovered that histone proteins form reversible DNA-protein cross-links (DPCs) with MdG in reconstituted NCPs, as well as in methylmethanesulfonate (MMS) treated cells. In this study, the formation and reactivity of MdG in MMS treated NCPs was examined at single nucleotide resolution. Sequences consisting of three or more consecutive dGs are more reactive with MMS. The efficiency and selectivity of MdG formation by MMS is largely unaffected within a NCP, although reactivity at several dGs is ∼1.5-2.5-fold higher in NCPs. DPC formation from MdG (DPC) predominates over AP at all positions within the NCP. With few exceptions, DPC yield is strongly dependent upon the accessibility of the major groove containing MdG to lysine-rich histone N-terminal tails. These data indicate that histone-MdG DPC formation will depend upon DNA sequence and translational position within an NCP.
Topics: DNA; DNA-Binding Proteins; Deoxyguanosine; Methyl Methanesulfonate; Models, Molecular; Molecular Structure; Nucleosomes; Particle Size; Surface Properties
PubMed: 31532638
DOI: 10.1021/acs.chemrestox.9b00314 -
Oxidative Medicine and Cellular... 2018Statins are 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, and this class of drugs has been studied as protective agents against DNA damages....
Statins are 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, and this class of drugs has been studied as protective agents against DNA damages. Alkylating agents (AAs) are able to induce alkylation in macromolecules, causing DNA damage, as DNA methylation. Our objective was to evaluate atorvastatin (AVA) antimutagenic, cytoprotective, and antigenotoxic potentials against DNA lesions caused by AA. AVA chemopreventive ability was evaluated using antimutagenicity assays (/microsome assay), cytotoxicity, cell cycle, and genotoxicity assays in HepG2 cells. The cells were cotreated with AVA and the AA methyl methanesulfonate (MMS) or cyclophosphamide (CPA). Our datum showed that AVA reduces the alkylation-mediated DNA damage in different experimental models. Cytoprotection of AVA at low doses (0.1-1.0 M) was observed after 24 h of cotreatment with MMS or CPA at their LC, causing an increase in HepG2 survival rates. After all, AVA at 10 M and 25 M had decreased effect in micronucleus formation in HepG2 cells and restored cell cycle alterations induced by MMS and CPA. This study supports the hypothesis that statins can be chemopreventive agents, acting as antimutagenic, antigenotoxic, and cytoprotective components, specifically against alkylating agents of DNA.
Topics: Alkylating Agents; Alkylation; Atorvastatin; Cell Cycle Checkpoints; Cell Nucleus; Cyclophosphamide; DNA Damage; Down-Regulation; Hep G2 Cells; Humans; Methyl Methanesulfonate; Salmonella enterica
PubMed: 29849914
DOI: 10.1155/2018/7820890 -
Molecular Microbiology Oct 2023Saccharomyces cerevisiae Pso2/SNM1 is essential for DNA interstrand crosslink (ICL) repair; however, its mechanism of action remains incompletely understood. While...
Saccharomyces cerevisiae Pso2/SNM1 is essential for DNA interstrand crosslink (ICL) repair; however, its mechanism of action remains incompletely understood. While recent work has revealed that Pso2/Snm1 is dual-localized in the nucleus and mitochondria, it remains unclear whether cell-intrinsic and -extrinsic factors regulate its subcellular localization and function. Herein, we show that Pso2 undergoes ubiquitination and phosphorylation, but not SUMOylation, in unstressed cells. Unexpectedly, we found that methyl methanesulfonate (MMS), rather than ICL-forming agents, induced robust SUMOylation of Pso2 on two conserved residues, K97 and K575, and that SUMOylation markedly increased its abundance in the mitochondria. Reciprocally, SUMOylation had no discernible impact on Pso2 translocation to the nucleus, despite the presence of steady-state levels of SUMOylated Pso2 across the cell cycle. Furthermore, substitution of the invariant residues K97 and K575 by arginine in the Pso2 SUMO consensus motifs severely impaired SUMOylation and abolished its translocation to the mitochondria of MMS-treated wild type cells, but not in unstressed cells. We demonstrate that whilst Siz1 and Siz2 SUMO E3 ligases catalyze Pso2 SUMOylation, the former plays a dominant role. Notably, we found that the phenotypic characteristics of the SUMOylation-defective mutant Pso2 closely mirrored those observed in the Pso2Δ petite mutant. Additionally, leveraging next-generation sequencing analysis, we demonstrate that Pso2 mitigates MMS-induced damage to mitochondrial DNA (mtDNA). Viewed together, our work offers previously unknown insights into the link between genotoxic stress-induced SUMOylation of Pso2 and its preferential targeting to the mitochondria, as well as its role in attenuating MMS-induced mtDNA damage.
Topics: Humans; Saccharomyces cerevisiae; Methyl Methanesulfonate; DNA, Mitochondrial; Sumoylation; Saccharomyces cerevisiae Proteins; Endodeoxyribonucleases; DNA Damage; Mitochondria; Translocation, Genetic; Ubiquitin-Protein Ligases
PubMed: 37649278
DOI: 10.1111/mmi.15145 -
Postepy Biochemii 1995
Review
Topics: Alkylating Agents; DNA Damage; DNA Repair; DNA Replication; DNA, Bacterial; Escherichia coli; Humans; Methyl Methanesulfonate; Methyltransferases; Mutagens; Neoplasms; O(6)-Methylguanine-DNA Methyltransferase
PubMed: 8797228
DOI: No ID Found -
Carcinogenesis Oct 1993Chinese hamster ovary CHO-B11 cells were exposed to methyl methanesulfonate (MMS) and the formation and repair of N-methylpurines were measured in the endogenous...
Chinese hamster ovary CHO-B11 cells were exposed to methyl methanesulfonate (MMS) and the formation and repair of N-methylpurines were measured in the endogenous dihydrofolate reductase (DHFR) gene domain and in mitochondrial DNA by alkaline hydrolysis which generates strand breaks at apurinic (AP) sites formed after neutral depurination. The initial levels of damage in the transcriptionally active DHFR gene, 3'-flanking non-transcribed region and in mitochondrial DNA were slightly different; the highest level of damage was in mitochondrial DNA, the lowest in the DHFR gene. The rate of adduct removal was similar in all three DNA regions examined. We conclude that there is no preferential repair of MMS induced N-methylpurines under these conditions, and that these lesions are efficiently removed from mitochondrial DNA.
Topics: Animals; CHO Cells; Cricetinae; DNA Repair; DNA, Mitochondrial; Methyl Methanesulfonate; Tetrahydrofolate Dehydrogenase
PubMed: 8222061
DOI: 10.1093/carcin/14.10.2105 -
Biology Letters Feb 2016Sea urchins are noted for the absence of neoplastic disease and represent a novel model to investigate cellular and systemic cancer protection mechanisms. Following...
Sea urchins are noted for the absence of neoplastic disease and represent a novel model to investigate cellular and systemic cancer protection mechanisms. Following intracoelomic injection of the DNA alkylating agent methyl methanesulfonate, DNA damage was detected in sea urchin cells and tissues (coelomocytes, muscle, oesophagus, ampullae and gonad) by the alkaline unwinding, fast micromethod. Gene expression analyses of the coelomocytes indicated upregulation of innate immune markers, including genes involved in NF-κB signalling. Results suggest that activation of the innate immune system following DNA damage may contribute to the naturally occurring resistance to neoplastic disease observed in sea urchins.
Topics: Animals; DNA Damage; Gene Expression; Immune System; Lytechinus; Methyl Methanesulfonate; Mutagens
PubMed: 26911343
DOI: 10.1098/rsbl.2015.1057 -
Journal of Chromatography. A Aug 2004A capillary gas chromatographic method using flame ionization detection was developed and validated for the trace analysis (ppm level) of methyl methanesulfonate, ethyl...
Trace analysis of residual methyl methanesulfonate, ethyl methanesulfonate and isopropyl methanesulfonate in pharmaceuticals by capillary gas chromatography with flame ionization detection.
A capillary gas chromatographic method using flame ionization detection was developed and validated for the trace analysis (ppm level) of methyl methanesulfonate, ethyl methanesulfonate, and isopropyl methanesulfonate in pharmaceutical drug substance. The method utilizes a megabore capillary column with bonded and crosslinked polyethylene glycol stationary phase. A dissolve-and-injection approach was adopted for sample introduction in a splitless mode. The investigated sample solvents include acetonitrile, ethyl acetate, methylene chloride, 1,2-dichloromethane, and toluene. Aqueous mixtures of acetonitrile and water can also be used as sample solvent. A limit of detection of about 1 microg/g (1 ppm) and limit of quantitation of 5 microg/g (5 ppm) were achieved for the mesylate esters in drug substance samples. The method optimization and validation are also discussed in this paper.
Topics: Chromatography, Gas; Ethyl Methanesulfonate; Mesylates; Methyl Methanesulfonate; Pharmaceutical Preparations; Reproducibility of Results; Sensitivity and Specificity
PubMed: 15387202
DOI: 10.1016/j.chroma.2004.06.095 -
Mutation Research Apr 1992The combined effects of methyl methanesulfonate (MMS) and ethyl methanesulfonate (EMS) on the induction of 6-thioguanine (6TG)-resistant mutants and chromosome...
The combined effects of methyl methanesulfonate (MMS) and ethyl methanesulfonate (EMS) on the induction of 6-thioguanine (6TG)-resistant mutants and chromosome aberrations were examined in Chinese hamster V79 cells. Cells were simultaneously treated with EMS at a concentration of D20 and MMS at various concentrations for 3, 6 or 9 h. In other experiments cells were simultaneously treated with MMS at a concentration of D20 and EMS at various concentrations for 3, 6 or 9 h. The mathematical analysis of the combined effects of both chemicals for cell killing (cytotoxicity) and 6TG-resistant mutations indicates that synergistic interactions were observed for both cell killing and mutations induced by MMS and EMS. The frequency of chromosome aberrations induced by simultaneous treatment with MMS at a concentration of D20 and EMS at various concentrations for 3 h was additive. However, the frequency of chromosome aberrations induced by EMS at a concentration of D20 and MMS at various concentrations for 3 h was not significantly different from those induced by MMS alone.
Topics: Animals; Cells, Cultured; Chromosome Aberrations; Cricetinae; Cricetulus; Drug Synergism; Ethyl Methanesulfonate; Methyl Methanesulfonate; Mutagens
PubMed: 1373826
DOI: 10.1016/0027-5107(92)90184-4