Journal: Cancer cell

Article Title: Small-molecule targeting of oncogenic FTO demethylase in acute myeloid leukemia

doi: 10.1016/j.ccell.2019.03.006

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: NB4 AML cells were incubated with two dosages of FB23-2, and the known histone demethylase inhibitors JIB-04 (HY-13953, MCE) ( Wang et al., 2013 ), Ciclopirox (T1482, Topscience) ( Yang et al., 2017 ), and IOX-1 (T6545, Topscience) ( Schiller et al., 2014 ) for 48 hr, respectively.

Techniques: Recombinant, Giemsa Stain, Staining, Proliferation Assay, Screening Assay, Isolation

Journal: mBio

Article Title: Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in Streptomyces coelicolor M145

doi: 10.1128/mBio.00326-19

Figure Lengend Snippet: Effect of ethanolamine (25 mM) as a sole nitrogen or carbon source on the biomass accumulation of the M145 parental strain, glnA4 mutant, epuRI mutant, and putative ethanolamine permease overexpression strain M145pRM4 spri_5940 in defined Evans medium after 7 days of incubation at 30°C. Error bars indicate standard errors of results from n = 3 biological replicates.

Article Snippet: Ethanolamine was derivatized using precolumn derivatization with ortho -phthalaldehyde (OPA)/mercaptoethanol (MCE) (Dr. Maisch GmbH, Ammerbuch, Germany).

Techniques: Mutagenesis, Over Expression, Incubation

Journal: mBio

Article Title: Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in Streptomyces coelicolor M145

doi: 10.1128/mBio.00326-19

Figure Lengend Snippet: Identification and organization of the novel ethanolamine utilization gene cluster in representative Streptomyces sp. genomes ( S. coelicolor , S. venezuelae , S. scabies , S. albus , and S. pristinaespiralis ). The proposed gene product functions are as follows: orf1 , membrane protein; orf2 , short-chain dehydrogenase; orf3 , aldehyde dehydrogenase; glnA4 , gamma-glutamylethanolamide synthetase; etuRI , ethanolamine utilization pathway regulator; orf4 , putative ethanolamine transporter; orf5 , glutamine amidotransferase.

Article Snippet: Ethanolamine was derivatized using precolumn derivatization with ortho -phthalaldehyde (OPA)/mercaptoethanol (MCE) (Dr. Maisch GmbH, Ammerbuch, Germany).

Techniques: Membrane

Journal: mBio

Article Title: Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in Streptomyces coelicolor M145

doi: 10.1128/mBio.00326-19

Figure Lengend Snippet: Physiological role of the glnA4 gene product in S. coelicolor M145 cells grown in the presence of ethanolamine. Phenotypic analysis of S. coelicolor M145 (A), the Δ glnA4 mutant (B), and the complemented mutant Δ glnA4 pRM4 glnA4 (C) was performed on defined Evans medium with ethanolamine hydrochloride (25 mM) (A) as the sole nitrogen source. Deletion of the glnA4 gene resulted in compromised growth on ethanolamine, whereas complementation with pRMglnA4 under the control of its native promoter restored growth on ethanolamine.

Article Snippet: Ethanolamine was derivatized using precolumn derivatization with ortho -phthalaldehyde (OPA)/mercaptoethanol (MCE) (Dr. Maisch GmbH, Ammerbuch, Germany).

Techniques: Mutagenesis, Control

Journal: mBio

Article Title: Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in Streptomyces coelicolor M145

doi: 10.1128/mBio.00326-19

Figure Lengend Snippet: Effect of glnA4 deletion on cell morphology in the presence of ethanolamine. The S. coelicolor M145 parental strain (A and B) and ΔglnA4 mutant (C and D) were cultivated in defined Evans medium with ethanolamine hydrochloride (EA; 25 mM) (left panels) or ammonium chloride (AM; 25 mM) (right panels). Phase-contrast microscopic pictures of the M145 parental strain and ΔglnA4 mutant mycelium were taken (under ×400 magnification) after 96 h of growth. Bar = 5 μm.

Article Snippet: Ethanolamine was derivatized using precolumn derivatization with ortho -phthalaldehyde (OPA)/mercaptoethanol (MCE) (Dr. Maisch GmbH, Ammerbuch, Germany).

Techniques: Mutagenesis

Journal: mBio

Article Title: Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in Streptomyces coelicolor M145

doi: 10.1128/mBio.00326-19

Figure Lengend Snippet: HPLC-based detection of ethanolamine in the supernatant from the 24-h and 96-h cultures of the M145 parental strain and ΔglnA4 mutant. Both strains were grown in defined Evans medium supplemented with ethanolamine (25 mM) as the sole N source. The HPLC chromatogram data are presented as follows: A, ethanolamine standard; B and C, detection of ethanolamine remaining in the supernatant of the M145 culture after 24 h (B) and 96 h (C) of growth; D and E, detection of ethanolamine remaining in the supernatant of the Δ glnA4 mutant culture after 24 h (D) and 96 h (E). The results indicate that only the S. coelicolor M145 parental strain was able to utilize more than one-third of ethanolamine from the medium after 96 h of incubation. mAU, milli-absorbance units.

Article Snippet: Ethanolamine was derivatized using precolumn derivatization with ortho -phthalaldehyde (OPA)/mercaptoethanol (MCE) (Dr. Maisch GmbH, Ammerbuch, Germany).

Techniques: Mutagenesis, Incubation

Journal: mBio

Article Title: Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in Streptomyces coelicolor M145

doi: 10.1128/mBio.00326-19

Figure Lengend Snippet: Transcriptional analysis of glnA4 in the presence of ammonium, polyamines, and ethanolamine as sole nitrogen sources. Data represent results of reverse transcriptase PCR analysis of glnA4 and hrdB (control) from S. coelicolor M145 cultivated in defined Evans medium with 5 mM (Am-) or 50 mM (Am+) concentrations of ammonium chloride, polyamines (Put, putrescine; Cad, cadaverine; Spd, spermidine, 25 mM each) or ethanolamine (EtAm; 25 mM) and glucose as the sole carbon source at a high concentration (Glc+; 25 g/liter) or a low concentration (Glc-; 2.5 g/liter). Total RNA was isolated from mycelium harvested after 24 h of cultivation in defined Evans medium.

Article Snippet: Ethanolamine was derivatized using precolumn derivatization with ortho -phthalaldehyde (OPA)/mercaptoethanol (MCE) (Dr. Maisch GmbH, Ammerbuch, Germany).

Techniques: Reverse Transcription, Control, Concentration Assay, Isolation

Journal: mBio

Article Title: Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in Streptomyces coelicolor M145

doi: 10.1128/mBio.00326-19

Figure Lengend Snippet: Effect of different amino group-containing compounds on the activity of GlnA4. All potential substrates of GlnA4 were present at a concentration of 50 mM. Mean values of n = 3 measurements are shown with standard deviations. GlnA4 is specific for ethanolamine.

Article Snippet: Ethanolamine was derivatized using precolumn derivatization with ortho -phthalaldehyde (OPA)/mercaptoethanol (MCE) (Dr. Maisch GmbH, Ammerbuch, Germany).

Techniques: Activity Assay, Concentration Assay