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Methyl methanesulfonate mms

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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.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (6 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Hydrogen peroxide h2o2, by Merck Group (4 mentions) Olaparib, by Selleck Chemicals (3 mentions) Propidium iodide, by Thermo Fisher Scientific (2 mentions) Sodium azide nan3, by Merck Group (2 mentions) Etoposide, by Merck Group (2 mentions) Talazoparib, by Selleck Chemicals (2 mentions) Veliparib, by Selleck Chemicals (2 mentions) Aphidicolin, by Merck Group (2 mentions) Resazurin, by Merck Group (2 mentions) Camptothecin, by Merck Group (2 mentions) Triton x 100, by Merck Group (2 mentions) Colchicine, by Merck Group (2 mentions) Benzo a pyrene, by Merck Group (2 mentions) Giemsa, by Merck Group (2 mentions) Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) High melting point agarose, by Merck Group (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Yeast extract, by BD (1 mentions)

49 protocols using methyl methanesulfonate mms

1

Evaluation of Cell Culture Reagents

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Dulbecco's modified Eagle's medium (DMEM), low-melting-point agarose (LMP), high-melting-point agarose (HMP), phosphate-buffered saline (PBS), and hydrogen peroxide (H2O2), amino acids, and nitrogen bases were purchased from Sigma (St. Louis, MO, USA). Foetal bovine serum (FBS) and penicillin/streptomycin were obtained from Gibco-BRL (Grand Island, NY, USA). MMS (methyl methanesulfonate) was purchased from Sigma (St. Louis, MO, USA). Yeast extract, bacto-peptone, bacto-agar, and yeast nitrogen base were obtained from Difco Laboratories (Detroit, MI). All other chemicals were of the highest purity grade commercially available.
Viau C.M., Moura D.J., Pflüger P., Facundo V.A, & Saffi J. (2016). Structural Aspects of Antioxidant and Genotoxic Activities of Two Flavonoids Obtained from Ethanolic Extract of Combretum leprosum. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 9849134.
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2

Culturing MCF-7 and MIA PaCa-2 Cell Lines

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Cell culture lines. Human breast cancer line, MCF-7 (7 (ATCC® HTB-22TM, breast adenocarcinoma)) and pancreatic cancer cell line MIA PaCa-2 (ATCC® CRL-1420TM, pancreas carcinoma) were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA). MCF-7 was maintained in RPMI supplemented with 10% FBS, non-EAA and sodium pyruvate. MIA PaCa-2 was maintained in DMEM supplemented with 10% FBS, 2.5% FHS and 1% penicillin/streptomycin. Both cells were cultured in a humidified 37 °C incubator with 5% CO2.
Cell culture reagents. RPMI Media 1640 and Dulbecco’s Modified Eagle’s Medium (DMEM) were purchased from Biowest. Fetal bovine serum (FBS), non-EAA, sodium pyruvate, 2.5% FHS, penicillin–streptomycin and TrypLE Express were purchased from Gibco.
General Reagents. MMS (methyl methanesulfonate) was purchased from Sigma-Aldrich; PBS (phosphate buffered saline) from Lonza; DMSO from Merck; propidium iodide from Invitrogen Molecular Probes; Hoechst 33342 solution from Fischer Scientific; calcein-AM from BD Biosciences (Franklin Lakes, NJ, USA).
Mackenzie T.A., Reyes F., Martínez M., González-Menéndez V., Sánchez I., Genilloud O., Tormo J.R, & Ramos M.C. (2024). Naphthoquinone Derivatives from Angustimassarina populi CF-097565 Display Anti-Tumour Activity in 3D Cultures of Breast Cancer Cells. Molecules, 29(2), 425.
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3

DNA Damage Response Protein Phosphorylation

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The antibodies used in this work include the following commercial antibodies: anti-phospho-histone H2AX (Ser139) (#80312-mouse), anti-phospho-53BP1 (Ser1778) (#2675-rabbit), anti-phospho-ATR (Thr1989) (#30632-rabbit), anti-phospho-Rad50 (Ser635) (#14223-rabbit) and anti-GAPDH (#3683-rabbit-HRP conjugated) from Cell Signaling Technology, USA. Other commercial antibodies include anti-phospho-(Ser/Thr) ATM/ATR Substrate (#AP0933-rabbit), anti-phospho-PRKDC (Ser2056) (#AP0621-rabbit) and anti-phospho-ATM (Ser1981) (#AP0008 -rabbit) from ABclonal, USA. Methyl methanesulfonate (MMS, Sigma, USA), DNA-PK inhibitor (AZD7648, AstraZeneca, UK), generously provided by Dr. Jennifer A. Black (Dept of Cellular and Molecular Biology and Pathogenic Bioagents, Ribeirão Preto Medical School, USP).
Gonzáles-Córdova R.A., dos Santos T.R., Gachet-Castro C., Andrade Vieira J., Trajano-Silva L.A., Sakamoto-Hojo E.T, & Baqui M.M. (2024). Trypanosoma cruzi infection induces DNA double-strand breaks and activates DNA damage response pathway in host epithelial cells. Scientific Reports, 14, 5225.
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4

Synthesis and Use of IQG-607 Compound

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The pentacyano(isoniazid)ferrate(II) compound (IQG-607) (S1 Fig) was synthesized according to Oliveira et al., (2006) [19 (link)]. For all treatments, IQG-607 was dissolved in saline solution (0.9% NaCl) immediately prior to use. Amphotericin B (AMB; Sigma-Aldrich) was used as a positive control to inhibit L. braziliensis proliferation. Methyl methanesulfonate (MMS; Sigma-Aldrich) or dimethyl sulfoxide (DMSO, Sigma-Aldrich) were used as positive controls in toxicity assays.
Amorim C.F., Galina L., Carvalho N.B., Sperotto N.D., Pissinate K., Machado P., Campos M.M., Basso L.A., Rodrigues-Junior V.S., Carvalho E.M, & Santos D.S. (2017). Inhibitory activity of pentacyano(isoniazid)ferrate(II), IQG-607, against promastigotes and amastigotes forms of Leishmania braziliensis. PLoS ONE, 12(12), e0190294.
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5

Gibson Assembly Reagents Preparation

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Enzymes for Gibson Assembly preparation were purchased in NEB (Ipswich, MA, USA). Media were from Helicon (Moscow, Russia). Oligonucleotides were made by Syntol (Moscow, Russia). All chemicals were of analytical purity. Hydrogen peroxide (H2O2) was obtained from Ferraine (Moscow, Russia). Mitomycin C (MitC) and methyl methanesulfonate (MMS) were obtained from Sigma-Aldrich (St. Louis, MO, USA). 2,2′-bis(bicyclo[2.2.1] heptane) (BBH) was synthetized as in previous work [32 (link)]. All test solutions and their dilutions were prepared immediately before use.
Kessenikh A.G., Novoyatlova U.S., Bazhenov S.V., Stepanova E.A., Khrulnova S.A., Gnuchikh E.Y., Kotova V.Y., Kudryavtseva A.A., Bermeshev M.V, & Manukhov I.V. (2021). Constructing of Bacillus subtilis-Based Lux-Biosensors with the Use of Stress-Inducible Promoters. International Journal of Molecular Sciences, 22(17), 9571.
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6

Generation and Characterization of Engineered Cell Lines

Cited 2 times
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All cell lines were obtained from American Type Culture Collection (Manassas, VA) and cultured in the recommended media in a humidified 5% CO2 incubator at 37°C. To generate MDA231-Alk5TD, -Alk5KR, and -vec, retroviruses encoding TβRI(Alk5)T204D, Alk5K232R (3 (link)), or the empty pBMN-I-GFP vector were produced by transfecting Ampho-Phoenix cells and then utilized for transduction, followed by green fluorescent protein (GFP) selection. The miR-181a/b and MSH2 expression plasmids were constructed and described elsewhere (18 (link), 19 (link)). The BRCA1 expression construct was kindly provided by Dr. Jeffrey D. Parvin (Ohio State University). The ATM expression construct (23 (link)) was obtained from Addgene (Cambridge, MA). Plasmid constructions and additional reagents are described in Supplementary Material. Cell transfection, reporter assays, production of viruses, as well as infection and selection of transduced cells were carried out as previously described (19 (link)). Recombinant human TGFβ1 was purchased from R&D Systems (Minneapolis, MN). The type I/II TGFβ receptor inhibitor LY2109761 was provided by Eli Lilly and Company (Indianapolis, IN). ABT-888 was purchased from ChemieTek (Indianapolis, IN). 4-Amino-1,8-naphthalimide (ANI), doxorubicin, methyl methanesulfonate (MMS), and 6-thioguanine (6-TG) were purchased from Sigma (St. Louis, MO).
Liu L., Zhou W., Cheng C.T., Ren X., Somlo G., Fong M.Y., Chin A.R., Li H., Yu Y., Xu Y., O'Connor S.T., O'Connor T.R., Ann D.K., Stark J.M, & Wang S.E. (2014). TGFβ Induces ‘BRCAness’ and Sensitivity to PARP Inhibition in Breast Cancer by Regulating DNA Repair Genes. Molecular cancer research : MCR, 12(11), 1597-1609.
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7

Candida Strains and Cell Line Maintenance

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Candida strains (listed in S1 Table) were provided by the Dr. P. David Rogers lab of the University of Tennessee Health Science Center, Memphis, TN. Human cell lines; THP-1 (ATCC, TIB-202), HepG2 (ATCC, HB-8065), and A549 (ATCC, CCL-185) were purchased from the American Type Culture Collection (Manassas, VA, USA). Dulbecco′s Modified Eagle′s culture medium (DMEM), 1X trypsin-EDTA solution, fetal bovine serum (FBS), 100X penicillin/streptomycin solution, Amphotericin B, caspofungin, fluconazole, itraconazole, 5-fluorocytosine, YPD agar and broth, cytochalasin D, 3-(N-morpholino) propanesulfonic acid (MOPS) buffer, and methyl methanesulfonate (MMS) were all purchased form Sigma-Aldrich (St. Louis, MO, USA). RPMI-1640 medium was purchased from Corning Incorporated (Coring, NY, USA). Geneticin selective antibiotic (G-148 sulfate), phosphate buffered saline (PBS), rhodamine phalloidin, propidium iodide, PrestoBlue, and RNaseA enzyme were purchased from Life Technologies Corporation (Carlsbad, CA, USA). 4% Paraformaldehyde was purchased from Alfa Aesar (Ward Hill, MA, USA) and zymolase 20T was purchased from MP Biomedicals, LLC (Solon, OH, USA). The yeast deletion collections (~1,056 heterozygous mutants and ~4,320 homozygous mutants) were purchased from GE Healthcare Life Sciences (Pittsburg, PA, USA) and ThermoFisher Scientific (Waltham, MA, USA), respectively.
Alqahtani F.M., Arivett B.A., Taylor Z.E., Handy S.T., Farone A.L, & Farone M.B. (2019). Chemogenomic profiling to understand the antifungal action of a bioactive aurone compound. PLoS ONE, 14(12), e0226068.
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8

MMS-Induced DNA Damage Sensitivity Assay

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W303a, NFY24, NFY26, and NFY27 strains were tested for MMS-induced DNA damage sensitivity. All strains were grown in YPD medium overnight at 30°C. The next day, secondary culture was grown until 0.5 OD600 at 30°C. After OD600 reached to 0.5, the culture was divided into two sets. One set of cells was treated with 0.03% (vol/vol) of methyl-methane-sulfonate (MMS) (Sigma Aldrich) and grown at 30°C for 2 h and another set was continuously grown at 30°C for 2 h without MMS. After that, the cells were washed twice and serially diluted, and 1000 cells of each culture were spread on respective selective media or YPD plates. The plates were incubated at 30°C for 40 h and the colonies obtained were counted in both treated and untreated conditions. Subsequently, the percentage survivability was calculated using the following formula: % survivability = [(number of cells grown on MMS plate)/(number of cells grown on untreated plate)] *100.
Fangaria N., Rani K., Singh P., Dey S., Kumar K.A, & Bhattacharyya S. (2022). DNA damage-induced nuclear import of HSP90α is promoted by Aha1. Molecular Biology of the Cell, 33(14), ar140.
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9

Genotoxicity Evaluation of Chemicals

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Sodium azide (NaN3) (CAS No. 26628-22-8), methylmethanesulfonate (MMS) (CAS No. 66-27-3), 2-aminoanthracene (2-AA) (CAS No. 613-13-8), cyclophosphamide (CPA) (CAS No. 50-18-0), and colchicine (CAS No. 64-86-8) were obtained from Sigma-Aldrich. 4-nitro-o-phenylene-diamine (4-NOPD) (CAS No. 99-56-9) was from Fluka. These chemicals were used as positive controls in the genotoxicity studies.
Dybdahl M., Selesko D.B, & Mikkelsen U.R. (2021). Safety evaluation of whey derived beta-lactoglobulin, Lacprodan® BLG. Toxicology Reports, 8, 617-626.
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10

Modulatory Effect of Betulinic Acid on DNA Damage

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The following four mutagens with different mechanisms of action were used to study the modulatory effect of BA on DNA damage: (i) methyl methanesulfonate (MMS; Sigma-Aldrich) dissolved in phosphate-buffered saline (PBS) at a concentration of 400 μM [11 (link)]. MMS is a direct-acting monofunctional alkylating agent that reacts with the DNA molecule by transferring methyl radicals [12 (link)]; (ii) doxorubicin (DXR; Eurofarma Laboratórios Ltda.) dissolved in sterile distilled water and used at a concentration of 0.3 μM [13 (link)]. DXR, one of the most potent broad-spectrum antitumor anthracycline antibiotics, is a free radical generator and a potent inhibitor of topoisomerase II [14 (link)]; (iii) (S)-(+)-camptothecin (CPT; Sigma-Aldrich) dissolved in DMSO (Sigma-Aldrich) at a concentration of 123.4 μM CPT acts by inhibiting topoisomerase I, an enzyme necessary for DNA replication [15 (link)]; (iv) etoposide (VP-16; Sigma-Aldrich) dissolved in DMSO at a concentration of 1.7 μM. VP-16 is a potent anticancer agent that inhibits topoisomerase II [16 (link)].
Acésio N.O., de Oliveira P.F., Mastrocola D.F., Lima I.M., Munari C.C., Sato V.L., Souza A.A., Flauzino L.G., Cunha W.R, & Tavares D.C. (2016). Modulatory Effect of Betulinic Acid on the Genotoxicity Induced by Different Mutagens in V79 Cells. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 8942730.
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