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Methyl Methanesulfonate

Methyl Methanesulfonate (MMS) is a chemical compound commonly used in biomedical research as a DNA alkylating agent.
It plays a key role in studying DNA damage, repair mechanisms, and mutagenesis.
MMS induces DNA lesions, such as 7-methylguanine and 3-methyladenine, which can lead to replication errors and genomic instability.
Researchers utilize MMS to investigate cellular responses to genotoxic stress, including cell cycle checkpoints, apoptosis, and DNA repair pathways.
Understanding the impact of MMS on biological systems provides insights into the underlying mechanisms of chemical mutagenesis and carcinogenesis.
This MeSH term description offers a concise overview of the key features and applications of Methyl Methanesulfonat1e in the context of biomedical research.

Most cited protocols related to «Methyl Methanesulfonate»

1
Comprehensive Stress Phenotype Analysis
For analysing growth phenotypes at distinct temperature, we monitored the growth of each mutant at a range of temperatures (25, 30, 37 and 39 °C) on agar-based yeast extract-peptone dextrose (YPD) medium. For analysing stress-related phenotypes and antifungal drug susceptibility, cells grown at 30 °C in liquid YPD medium for 16 h were 10-fold serially diluted (1 to 104 dilutions) and spotted on YPD medium containing the indicated concentrations of the following chemicals: osmotic (sorbitol) and cation/salt stresses (NaCl and KCl) under either glucose-rich (YPD) or glucose-starved (yeast extract-peptone, YP) conditions; oxidative stress (hydrogen peroxide (H2O2), tert-butyl hydroperoxide (an organic peroxide), menadione (a superoxide anion generator), diamide (a thiol-specific oxidant)); heavy-metal stress (CdSO4); genotoxic stress (methyl methanesulfonate and hydroxyurea); cell membrane/wall-destabilizing stress (SDS, calcofluor white and Congo red); ER stress (tunicamycin and DTT); and antifungal agents (fludioxonil, FCZ, AmpB and flucytosine). Cells were incubated at 30 °C and photographed for 2–5 days.
Jung K.W., Yang D.H., Maeng S., Lee K.T., So Y.S., Hong J., Choi J., Byun H.J., Kim H., Bang S., Song M.H., Lee J.W., Kim M.S., Kim S.Y., Ji J.H., Park G., Kwon H., Cha S., Meyers G.L., Wang L.L., Jang J., Janbon G., Adedoyin G., Kim T., Averette A.K., Heitman J., Cheong E., Lee Y.H., Lee Y.W, & Bahn Y.S. (2015). Systematic functional profiling of transcription factor networks in Cryptococcus neoformans. Nature Communications, 6, 6757.
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Publication 2015
Agar Antifungal Agents calcofluor white Cells Diamide Flucytosine fludioxonil Genotoxic Stress Glucose Hydroxyurea Metals, Heavy Methyl Methanesulfonate Osmosis Oxidants Oxidative Stress Peptones Peroxide, Hydrogen Peroxides Phenotype Plasma Membrane Saccharomyces cerevisiae Salt Stress Sodium Chloride Sorbitol Sulfhydryl Compounds Superoxides Susceptibility, Disease Technique, Dilution tert-Butylhydroperoxide Tunicamycin Vitamin K3
2
Transposon library construction and screening
Library construction was done as described (van Opijnen et al. 2009 (link); van Opijnen and Camilli 2010 ). Note that the magellan6 minitransposon we designed lacks transcriptional terminators, therefore allowing for read-through transcription, which explains why no relevant polar effects were observed by examining fitness of downstream genes (Supplemental Table S1). Additionally, the minitransposon contains stop codons in all three frames in either orientation when inserted into a coding sequence. In vitro selection experiments were done with six independently generated libraries each with a size of ∼8000 transposon insertion mutants covering 88% of nonessential genes. Growth conditions where the carbon source was varied consisted of semi-defined minimal media (SDMM) at pH 7.3 supplemented with 10 mM of one of the following carbon sources: glucose, fructose, mannose, galactose, N-acetylglucosamine (GlcNac), sialic acid, sucrose, maltose, cellobiose, or raffinose. Stress conditions consisted of SDMM with 10 mM glucose at pH 7.3 and one of the following stresses: Metal stress, 0.5 mM of 2,2′-Bipyridyl (Sigma-Aldrich); DNA damage, Methyl methanesulfonate 0.015% (MMS, Fluka); hydrogen peroxide exposure, H2O2 4.5 mM (Sigma-Aldrich); acidic pH stress, pH6; temperature stress, growth at 30°C; antibiotic exposure, norfloxacin 1.5 μg/mL (Sigma-Aldrich); and DNA transformation.
Nasopharynx colonization experiments were done in 17 mice with eight independently generated libraries each with a size of ∼4000 mutants, while lung infection experiments were done in 20 mice with six libraries each with a size of ∼30,000 mutants. Because of differences in the bacterial load, 105–106 colony forming units (cfu) for nasopharynx and 107–108 cfu for lung, smaller libraries were used for the nasopharynx in order to minimize the stochastic loss of mutants. Mice were euthanized after 24 h for lung infection, followed by removal and homogenization of the lungs, and 48 h for nasopharynx colonization, followed by flushing of the nasopharynx with 500 μL of PBS.
van Opijnen T, & Camilli A. (2012). A fine scale phenotype–genotype virulence map of a bacterial pathogen. Genome Research, 22(12), 2541-2551.
Publication 2012
Acetylglucosamine Acids Antibiotics Bipyridyl Carbon Carbon-10 Cellobiose Codon, Terminator DNA Damage DNA Library Fructose Galactose Genes Genetic Fitness Glucose Growth Disorders Infection Jumping Genes Lung Maltose Mannose Metals Methyl Methanesulfonate Mus N-Acetylneuraminic Acid Nasopharynx Norfloxacin Open Reading Frames Peroxide, Hydrogen Raffinose Reading Frames Stress Disorders, Traumatic Sucrose Transcription, Genetic
3
Systematic Phenotypic Profiling of Yeast Mutants
238 mutants, broadly spanning GO Biological Function categories plus several uncharacterised genes, were selected from a prototroph derivative of the Bioneer deletion library (Sideri et al., 2015 (link)). Strains were arranged in 384-colony (16 × 24) format with a single 96 grid placed in the top left position, so that the grid includes one colony in every fourth position within the 384-colony array. To prepare replicates, this plate was independently pinned 3 times from the cryostock on solid YES media for each batch. From these plates, colonies were then spotted on assay plates containing various toxins, drugs or nutrients. The conditions used in Figure 3B are: EtOH10 is YES+10% (v/v) ethanol, VPA10 is YES+10 mM valproic acid, MMS0.005 is YES+0.005% (v/v) methyl methanesulfonate, Forma2.5 is YES+2.5% (v/v) formamide, Diamide2 is YES+2 mM diamide, EMM is standard Edinburgh Minimal Medium, YES is standard Yeast extract with supplements and 3% glucose. Assay plates were usually grown for 2 days at 32°C but this varied according to the strength of the stress slowing the growth of the colonies. After incubation, images were acquired using EPSON V800 scanners and pyphe-scan and quantified with gitter (see options above) or pyphe-quantify in redness mode. Grid correction and subsequent row/column median normalisation of maximum slopes and individual timepoints was performed in pyphe-analyse. Row/column median normalisation was applied to redness data plates. For the size data set, 0-sized colonies and colonies with a circularity below 0.85 were set to NA. Plates with a CV > 0.2 or FUV >1 were removed as those most likely represent conditions in which the stress was too strong or where technical errors occurred.
Kamrad S., Rodríguez-López M., Cotobal C., Correia-Melo C., Ralser M, & Bähler J. (2020). Pyphe, a python toolbox for assessing microbial growth and cell viability in high-throughput colony screens. eLife, 9, e55160.
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Publication 2020
Biological Assay Biological Processes Deletion Mutation Diamide Dietary Supplements DNA Library Erythema Ethanol formamide Genes Glucose Methyl Methanesulfonate Nutrients Pharmaceutical Preparations Radionuclide Imaging Saccharomyces cerevisiae Strains Toxins, Biological Valproic Acid
4
Cell Lines and Reagents for Cancer Research
DU145, CCRF-CEM, MOLT4, and K562 were obtained from the Division of Cancer Treatment (DCTD), Developmental Therapeutics Program (DTP, NCI), and EW8 and A673 are kind gifts from Dr. Lee Helman (NCI/NIH). All cells were grown in RPMI medium with 10% FBS (Gibco-BRL) at 37°C in 5% CO2. Information about the SCLC lines is shown in Table S2. The ATR inhibitor VE-821, olaparib, and veliparib were obtained from the DCTD. Talazoparib was provided by BioMarin Pharmaceutical Inc. Temozolomide (T2577) and methyl methanesulfonate MMS (129925) were purchased from Sigma-Aldrich.
Murai J., Feng Y., Yu G.K., Ru Y., Tang S.W., Shen Y, & Pommier Y. (2016). Resistance to PARP inhibitors by SLFN11 inactivation can be overcome by ATR inhibition. Oncotarget, 7(47), 76534-76550.
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Publication 2016
Cells Gifts Malignant Neoplasms Methyl Methanesulfonate olaparib Pharmaceutical Preparations Program Development Small Cell Lung Carcinoma talazoparib Temozolomide Therapeutics VE 821 veliparib
5
Evaluating BRCA2 Variant Function
The selected missense variants were cloned into a 3x FLAG-tagged full-length BRCA2 cDNA expression plasmid using the QuickChange site-directed mutagenesis kit XL (Stratagene) (13 (link)). Mutations were verified by DNA sequencing. Wild-type and mutant constructs were co-transfected with an I-Sce1–expressing pcBASce plasmid (13 (link)) into V-C8 cells containing the DR-GFP reporter plasmid. The BRCA2 deficient V-C8 hamster lung fibroblast cell line (XRCC11) displays chromosomal instability and abnormal centrosomes, reduced nuclear localization of the RAD51 protein that is central in the HDR processes, and sensitivity to cross-linking agents such as methyl methanesulfonate (MMS) (19 (link)). Cells expressing green fluorescent protein (GFP) following HDR-dependent repair of an I-Sce1 induced DNA double strand break in the DR-GFP reporter plasmid were quantified by fluorescent activated cell sorting (FACS) after 72 hrs. Transfection efficiency was evaluated by immunofluorescence (IF) based counting of GFP positive cells. Expression of wildtype and mutant BRCA2 was evaluated by immunoblotting with rabbit Anti-Flag antibody (1:1000 F7425 Sigma) of mouse Anti-Flag M2 (F1804 Sigma) immunoprecipitates from protein lysates.
Guidugli L., Pankratz V.S., Singh N., Thompson J., Erding C.A., Engel C., Schmutzler R., Domchek S., Nathanson K., Radice P., Singer C., Tonin P.N., Lindor N.M., Goldgar D.E, & Couch F.J. (2012). A classification model for BRCA2 DNA binding domain missense variants based on homology directed repair activity. Cancer research, 73(1), 265-275.
Publication 2012
Antibodies, Anti-Idiotypic Cell Lines Cells Centrosome Chromosomal Instability DNA, Complementary DNA Breaks, Double-Stranded Fibroblasts Gene, BRCA2 Green Fluorescent Proteins Hamsters Hypersensitivity Immunofluorescence Lung Methyl Methanesulfonate Missense Mutation Mus Mutagenesis, Site-Directed Mutation Plasmids Proteins Rabbits Rad51 Recombinase Transfection

Most recents protocols related to «Methyl Methanesulfonate»

1
Genotoxic Stress Response in Yeast
Single colony derived yeast cells were incubated overnight at the appropriate temperature in liquid medium. Cells were diluted to 0.5 OD600 and spotted in ten-fold serial dilutions onto YPD plates, SC plates, or plates containing the indicated amount of genotoxic drugs, i.e., methylmethanesulfonate (MMS), camptothecin (CPT) or hydroxyurea (HU) (all drugs: Sigma-Aldrich). The agar plates were incubated at the indicated temperatures and time and imaged using the ChemiDoc™ Touch Imaging System (Bio-Rad).
Standard YPD agar has pH 5.5. To make YPD agar plates with alkaline pH, we titrated melted YPD agar with 10 N NaOH until pH 8.0 was reached. The wsc1::KAN knockout strain was used as a positive control for the alkaline agar plates28 (link).
Schindler N., Tonn M., Kellner V., Fung J.J., Lockhart A., Vydzhak O., Juretschke T., Möckel S., Beli P., Khmelinskii A, & Luke B. (2023). Genetic requirements for repair of lesions caused by single genomic ribonucleotides in S phase. Nature Communications, 14, 1227.
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Publication 2023
Agar Camptothecin Cells Hydroxyurea Methyl Methanesulfonate Pharmaceutical Preparations Strains Technique, Dilution Touch Yeasts
2
Analyzing DNA Damage and Repair in Cancer Cells
Immunofluorescence assay was performed for checking the BRCA1 and H2AX foci formation in cancer cells and in the cells which were treated with 0.035% MMS (methyl methane sulfonate) to check the repair activity using the following protocol: 5 × 104 cells were seeded on collagen-coated coverslips in each well of a six-well plate. At 80% confluency, the cells were washed with 1X PBS and fixed in 4% paraformaldehyde, followed by permeabilization with 0.3% triton-X 100 in 1X PBS for 10 min each. Blocking was performed for 1 h with 5% FBS in 1X PBS at room temperature. The cells were incubated with primary antibodies overnight at 4°C in humid conditions, followed by the corresponding secondary antibody for 1 h at room temperature. After each antibody treatment, the cells were washed three times with 1X PBS. The coverslips containing cells were mounted on a glass slide with 10 ul of PD-DAPI, and imaging was carried out using an EVOS-M5000 microscope. For checking the repair activity, 0.035% MMS was added and incubated at 37°C for 15 min. After 15 min, the MMS was washed off with 1X PBS 2–3 times and released in fresh recommended culture media. The cells were fixed at 0 and 24 h after recovery. The cells without MMS were maintained as a control.
Katheeja M.N., Das S.P., Das R, & Laha S. (2023). BRCA1 interactors, RAD50 and BRIP1, as prognostic markers for triple-negative breast cancer severity. Frontiers in Genetics, 14, 1035052.
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Publication 2023
Antibodies BRCA1 protein, human Collagen Culture Media DAPI Immunofluorescence Immunoglobulins Malignant Neoplasms Methyl Methanesulfonate Microscopy paraform Triton X-100
3
Rutin and Quercetin Compound Dilution
Aliquots of rutin; hydrolyzed rutins (4, 8, and 12 h reaction times); quercetin and methyl methanesulfonate (MMS, Sigma-Aldrich, Cod.78697) were diluted (1:10 p/v) in dimethyl sulfoxide (DMSO, Synth) before dilution in complete medium to afford the final concentrations described in each assay.
Sobreiro M.A., Della Torre A., de Araújo M.E., Canella P.R., de Carvalho J.E., Carvalho P.D, & Ruiz A.L. (2023). Enzymatic Hydrolysis of Rutin: Evaluation of Kinetic Parameters and Anti-Proliferative, Mutagenic and Anti-Mutagenic Effects. Life, 13(2), 549.
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Publication 2023
Biological Assay Methyl Methanesulfonate Quercetin Rutin Sulfoxide, Dimethyl Technique, Dilution
4
Yeast Stress Response Assay
Wt, cdh1Δ, yap1Δ and yap1Δcdh1Δ strains were grown overnight at 26 °C in YPGal medium until mid-log phase. Each culture was then diluted to OD600nm = 0.1 and ten-fold dilutions were performed using PBS buffer. Cells were spotted in YPGal plates, used within 48h of preparation, supplemented with 0, 2.5 and 5 mM of H2O2 (Merck, Darmstadt, Germany) and 0.05% (v/v) of methyl methanesulfonate (MMS, ThermoFisher Scientific, Waltham, MA, USA). Cells were incubated for 2 days at 26 °C.
Leite A.C., Barbedo M., Costa V, & Pereira C. (2023). The APC/C Activator Cdh1p Plays a Role in Mitochondrial Metabolic Remodelling in Yeast. International Journal of Molecular Sciences, 24(4), 4111.
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Publication 2023
Buffers Cells Methyl Methanesulfonate Peroxide, Hydrogen Strains Technique, Dilution
5
EC50 Determination of Compounds in Hep G2 Cells
The EC50 determinations of the isolated compounds were performed with 12-point dose-response curves. Each concentration was evaluated in triplicate (concentration range of 50–0.0244 µM) in an MTT-based assay performed in liver carcinoma Hep G2 HB-8065 ATCC (American Type Culture Collection) cells as previously described by Annang et al., 2020 [29 (link)]. Cells (96-well plates were used with seeding at 1 × 104 cells/well) were cultured in 200 µL MEM medium per well at 37 °C, 5% CO2 for 24 h, after which the spent media were replaced with 200 µL MEM medium and 1 μL of test compounds. DMSO (0.5%), 8 mM methyl methanesulfonate and doxorubicin were used as positive, negative, and standard drug controls, respectively. After 72 h incubation with the test compounds at 37 °C, 100 µL of 0.5 mg/mL MTT solution diluted in MEM without phenol red was added to each well, and plates were briefly shaken and further incubated for 3 h at 37 °C. The supernatant in each well was carefully discarded and replaced with 100 µL of DMSO, and plates were gently shaken (to solubilize the formazan formed) before reading the absorbance at 570 nm in a Victor2 plate reader (Perkin Elmer, USA).
Annang F.B., Pérez-Moreno G., Bosch-Navarrete C., González-Menéndez V., Martín J., Mackenzie T.A., Ramos M.C., Ruiz-Pérez L.M., Genilloud O., González-Pacanowska D., Vicente F, & Reyes F. (2023). Antiparasitic Meroterpenoids Isolated from Memnoniella dichroa CF-080171. Pharmaceutics, 15(2), 492.
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Publication 2023
Biological Assay Cells Doxorubicin Formazans Hepatocellular Carcinomas Methyl Methanesulfonate Sulfoxide, Dimethyl

Top products related to «Methyl Methanesulfonate»

1
Methyl methanesulfonate by Merck Group
Sourced in United States, Germany, France, Spain
Methyl methanesulfonate is a colorless, viscous liquid chemical compound. It is commonly used as a research tool in laboratory settings.
2
Methyl methanesulfonate mms by Merck Group
Sourced in United States, Israel
Methyl methanesulfonate (MMS) is a chemical compound used in laboratory settings. It is a colorless, oily liquid with a pungent odor. MMS is primarily used as a DNA alkylating agent in various research applications.
3
Dimethyl sulfoxide (dmso) by Merck Group
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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
4
Etoposide by Merck Group
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Etoposide is a chemotherapeutic agent used in the treatment of various types of cancer. It is a topoisomerase inhibitor that disrupts the process of DNA replication, leading to cell death. Etoposide is available as a solution for intravenous administration.
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Fetal bovine serum (fbs) by Thermo Fisher Scientific
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Hydroxyurea by Merck Group
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Hydroxyurea is a chemical compound used in various laboratory applications. It functions as an inhibitor of ribonucleotide reductase, an enzyme involved in the synthesis of DNA.
7
Cisplatin by Merck Group
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Cisplatin is a platinum-based medication used as a chemotherapeutic agent. It is a crystalline solid that can be dissolved in water or saline solution for administration. Cisplatin functions by interfering with DNA replication, leading to cell death in rapidly dividing cells.
8
Olaparib by Selleck Chemicals
Sourced in United States, Germany, United Kingdom, China, France, Japan
Olaparib is a laboratory chemical product. It is a poly(ADP-ribose) polymerase (PARP) inhibitor. Olaparib functions by inhibiting the activity of PARP enzymes, which are involved in DNA repair processes.
9
Triton x 100 by Merck Group
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Triton X-100 is a non-ionic surfactant commonly used in various laboratory applications. It functions as a detergent and solubilizing agent, facilitating the solubilization and extraction of proteins and other biomolecules from biological samples.
10
Camptothecin by Merck Group
Sourced in United States, Germany, United Kingdom, Belgium, Sao Tome and Principe, France, Macao, Poland
Camptothecin is a naturally occurring alkaloid compound isolated from the Camptotheca acuminata tree. It is a biologically active compound with potential applications in research and development. Camptothecin exhibits inhibitory effects on the enzyme topoisomerase I, which is involved in DNA replication and transcription processes. This property makes Camptothecin a valuable tool for studying cellular mechanisms and biological processes.

More about "Methyl Methanesulfonate"

Methyl Methanesulfonate (MMS) is a widely-used chemical compound in biomedical research, known for its role as a DNA alkylating agent.
This compound plays a crucial part in studying DNA damage, repair mechanisms, and mutagenesis.
MMS induces specific DNA lesions, such as 7-methylguanine and 3-methyladenine, which can lead to replication errors and genomic instability.
Researchers frequently utilize MMS to investigate cellular responses to genotoxic stress, including cell cycle checkpoints, apoptosis, and DNA repair pathways.
Understanding the impact of MMS on biological systems provides invaluable insights into the underlying mechanisms of chemical mutagenesis and carcinogenesis.
MMS is often used in conjunction with other compounds, like DMSO, Etoposide, Hydroxyurea, Cisplatin, Olaparib, and Camptothecin, to study various aspects of DNA damage and cellular responses.
Additionally, FBS and Triton X-100 may be used in MMS-based experiments to support cell culture and sample preparation.
By leveraging the synergies between these related compounds and techniques, researchers can gain a more comprehensive understanding of the complex interplay between DNA damage, cellular signaling, and genomic integrity.
The versatility and widespread use of Methyl Methanesulfonate (MMS) in biomedical research underscores its importance as a tool for advancing our knowledge of fundamental biological processes and mechanisms underlying genetic instability and disease.
Whether you're studying DNA repair, mutagenesis, or cellular stress responses, MMS can be a valuable component of your research toolkit.