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Journal of Biochemistry Aug 2006Complexes of actinomycin D (AMD) and 7-amino-actinomycin D (7AAMD) with model hairpin oligonucleotide HP1 and various types of DNA in aqueous solutions were investigated...
Complexes of actinomycin D (AMD) and 7-amino-actinomycin D (7AAMD) with model hairpin oligonucleotide HP1 and various types of DNA in aqueous solutions were investigated by steady-state, polarized, time-resolved and stopped-flow fluorimetry, and photometry. Prompt non-stacking binding of the actinomycins inside HP1 was observed. No energy transfer from nucleotides to 7AAMD in the complex was detected, most likely because of the absence of stacking intercalation. Complex formation of AMD or 7AAMD and HP1 was followed by the transition from a random flexible conformation of the hairpin to a more compact rigid structure, and subsequently to hypochromism. Strong competition between AMD and 7AAMD for a cavity in HP1 was observed. The decrease in the 7AAMD emission after addition of DNA to the 7AAMD/HP1 complex indicates that actinomycins can be redistributed from HP1 to DNA, i.e. hairpin oligonucleotides can serve as molecular carriers of actinomycins.
Topics: DNA; Dactinomycin; Energy Transfer; Fluorescence; Nucleic Acids; Oligonucleotides; Protein Binding
PubMed: 16861251
DOI: 10.1093/jb/mvj150 -
Journal of Natural Products Mar 2000Structure elucidation of five components of the actinomycin Z complex (Z(1)-Z(5)) isolated from Streptomyces fradiae is described. The components were separated by Si...
Structure elucidation of five components of the actinomycin Z complex (Z(1)-Z(5)) isolated from Streptomyces fradiae is described. The components were separated by Si gel column chromatography and TLC/PLC and analyzed by ESIMS, FABMS, LC-MS of derivatized hydrolysates, and 2D NMR techniques. This permitted determination of the complete structures of actinomycins Z(1)-Z(5). In Z(3) and Z(5,) site 1 of the beta-depsipeptide is occupied by the rare 4-chloro-L-threonine, an amino acid not previously found in an actinomycin. The structural variants of the actinomycin Z complex have the molecular architecture typical of other actinomycins but possess greater structural diversity resulting from the presence of several highly unusual amino acids. Actinomycins Z(3) and Z(5,) but not Z(1), were more potent than actinomycin D in cytotoxicity assays against three tumor cell lines.
Topics: Antibiotics, Antineoplastic; Dactinomycin; Drug Screening Assays, Antitumor; Humans; Molecular Structure; Spectrum Analysis; Streptomyces; Threonine; Tumor Cells, Cultured
PubMed: 10757717
DOI: 10.1021/np990416u -
Natural Product Research Jun 2019Antifungal bioassays led to the isolation of actinomycins D and A from TRM45540 collected from Norpo in Xinjiang, and these compounds were identified by nuclear...
Antifungal bioassays led to the isolation of actinomycins D and A from TRM45540 collected from Norpo in Xinjiang, and these compounds were identified by nuclear magnetic resonance spectroscopy. The antifungal activity of actinomycin D was higher than that of actinomycin A. Actinomycin D clearly inhibited the spore germination, hyphal growth and biomass accumulation of in a dose-dependent manner. Flow cytometric analysis with propidium iodide, total ergosterol measurement, cell leakage and scanning electron microscopy experiments demonstrated that the plasma membrane of this fungus was damaged by actinomycin D, resulting in swollen cells and cellular content leakage. Transmission electron microscopy revealed that parts of the plasma membrane infolded after being treated with actinomycin D. The antifungal activity of actinomycin D damaged the fungal plasma membrane of via a membrane-splitting mechanism, which provided new insights into the functional mechanism of actinomycin D.
Topics: Antifungal Agents; Cell Membrane; Dactinomycin; Dose-Response Relationship, Drug; Streptomyces; Verticillium
PubMed: 29382222
DOI: 10.1080/14786419.2018.1431630 -
Journal of Chromatography. B,... Oct 2003Actinomycin D is an anti-cancer drug commonly used in the treatment of paediatric malignancies such as Wilms' tumour, Ewing's sarcoma and rhabdomyosarcoma. Despite its...
Actinomycin D is an anti-cancer drug commonly used in the treatment of paediatric malignancies such as Wilms' tumour, Ewing's sarcoma and rhabdomyosarcoma. Despite its long history of clinical use, little is known about the pharmacokinetics of actinomycin D in humans, largely due to problems in developing an analytical assay with the required sensitivity to measure relevant clinical concentrations. As actinomycin D treatment in children with cancer is associated with veno-occlusive disease (VOD), and as the dose intensity of actinomycin D treatment has been defined as a significant risk factor for the development of this potentially life-threatening hepatic toxicity, pharmacokinetic studies of actinomycin D may be beneficial in optimizing treatment with this drug. In order to investigate this issue, we developed a sensitive liquid chromatography-mass spectrometry (LC-MS) method for the determination of actinomycin D in human plasma samples. Extraction of analytical samples was carried out with acetonitrile and analysis performed on an API 2000 LC/MS/MS using an internal standard of 7-aminoactinomycin D. A limit of quantitation of 1.0 ng/ml was determined, allowing the reliable measurement of actinomycin D in plasma samples obtained from patients receiving this drug clinically. The method demonstrated good reproducibility, over the calibration curve range of 1.0-100 ng/ml, with intra- and inter-assay precision CVs of 2.7-11.3 and 2.3-7.8%, respectively. Accuracy data showed relative errors of 2.0-16.4 and 10.4-15.2% for intra-assay (n=10) and inter-assay (n=7) experiments, respectively. Initial results of actinomycin D pharmacokinetics in paediatric patients are shown.
Topics: Antibiotics, Antineoplastic; Child; Chromatography, Liquid; Dactinomycin; Humans; Neoplasms; Reference Standards; Spectrometry, Mass, Electrospray Ionization
PubMed: 14522028
DOI: 10.1016/s1570-0232(03)00573-7 -
Biochemistry Aug 1998We have examined the role of DNA composition in the binding of actinomycin D to single-stranded DNA. By using the fluorescent analogue 7-aminoactinomycin D, we were able...
We have examined the role of DNA composition in the binding of actinomycin D to single-stranded DNA. By using the fluorescent analogue 7-aminoactinomycin D, we were able to monitor binding of the drug to ssDNA with single base changes distant from the 5'-TAGT-3' site previously determined to be a high-affinity site for actinomycin D binding (Wadkins et al. (1996) J. Mol. Biol. 262, 53-68). Our binding studies indicated that secondary structures in the ssDNA were likely to be responsible for binding the drug. A series of six low-melting DNA hairpins containing all or part of the 5'-TAGT-3' binding site were synthesized. The highest Tm observed for the melting of these hairpins was 34.2 +/- 0.3 degrees C, and it depended on the length of the stem region. These metastable hairpins were stabilized by 7-aminoactinomycin D, with the drug shifting the Tm for the drug-hairpin complex to approximately 45 degrees C. The hairpins showed very high affinity (Kd approximately 0.1 microM) for 7-aminoactinomycin D, with some dependence on stem length. Digestion of the hairpins in the presence and absence of drug using mung bean nuclease, which specifically interacts with the loop region of hairpin DNA, revealed that the stable hairpins (i) contain a number of non-Watson-Crick base pairs, and (ii) undergo a conformational change in the loop region upon binding 7-aminoactinomycin D. Our results suggest that stabilization of unusual hairpins by actinomycin D may be an important aspect of the potent transcription inhibition activity of this drug.
Topics: Base Composition; Base Sequence; Binding Sites; DNA, Single-Stranded; Dactinomycin; Fluorescent Dyes; Models, Molecular; Nucleic Acid Conformation; Single-Strand Specific DNA and RNA Endonucleases; Temperature; Thermodynamics
PubMed: 9718315
DOI: 10.1021/bi9809730 -
The Journal of Cell Biology Jan 1971Nucelolar morphology was studied by electron microscopy in control and actinomycin D-treated populations of Tetrahymena pyriformis (W) during the cultural growth cycle....
Nucelolar morphology was studied by electron microscopy in control and actinomycin D-treated populations of Tetrahymena pyriformis (W) during the cultural growth cycle. Nucleoli exhibit an "aging" cycle concomitant with the cultural growth cycle, but independent of the individual cell cycle. Four different stages in the course of this aging process have been defined. Stage 1 occurs upon inoculation (low number of cells per milliliter) and lasts through lag and accelerating growth phases. In this stage, many small nucleoli are found at the nuclear periphery. In stages 2 and 3, nucleolar fusion begins. Stage 2 dominates the first half of logarithmic growth, and stage 3 dominates the second half. In late decelerating growth phase, the nucleoli enter stage 4. In this stage, only a few large nucleoli are present and these are apparently inactive in ribosome production. In stationary phase, where total RNA remains constant, only stage 4 nucleoli are present. The relative preponderance of granular vs. fibrous components in the nucleoli changes during this cycle, the granular component dominating stage 1 nucleoli and the fibrillar, stage 4 nucleoli. There is a shortening of the intermediate nucleolar stages in the treated cultures; fusion occurs early and is now pronounced. Not enough ribosomes accumulate to carry the treated cultures through the number of generations equivalent to those of the control, which produces a premature stationary phase.
Topics: Animals; Cell Nucleolus; Culture Media; Dactinomycin
PubMed: 5545100
DOI: 10.1083/jcb.48.1.143 -
Journal of Virology Jan 1971The in vitro product of mouse leukemia virus deoxyribonucleic acid (DNA) polymerase can be separated into two fractions by sedimentation in sucrose gradients. These two...
The in vitro product of mouse leukemia virus deoxyribonucleic acid (DNA) polymerase can be separated into two fractions by sedimentation in sucrose gradients. These two fractions were analyzed for their content of single-stranded DNA, double-stranded DNA, and DNA-ribonucleic acid (RNA) hybrid by (i) digestion with enzymes of known specificity and (ii) equilibrium centrifugation in Cs(2)SO(4) gradients. The major fraction early in the reaction contained equal amounts of single-stranded DNA and DNA-RNA hybrid and little double-stranded DNA. The major fraction after extensive synthesis contained equal amounts of single-and double-stranded DNA and little hybrid. In the presence of actinomycin D, the predominant product was single-stranded DNA. To account for these various forms of DNA, we postulate the following model: the first DNA synthesis occurs in a replicative complex containing growing DNA molecules attached to an RNA molecule. Each DNA molecule is displaced as single-stranded DNA by the synthesis of the following DNA strand, and the single-stranded DNA is copied to form double-stranded DNA either before or after release of the single strand from the RNA. Actinomycin blocks this conversion of single-to double-stranded DNA.
Topics: Carbon Isotopes; Centrifugation, Density Gradient; Cesium; DNA Replication; Dactinomycin
PubMed: 5543423
DOI: 10.1128/JVI.7.1.106-111.1971 -
Bioorganicheskaia Khimiia 2014Variety of different compounds use for delivery of antibiotics to the tumor cells. In this work, using a highly sensitive fluorescence analysis, we have studied...
Variety of different compounds use for delivery of antibiotics to the tumor cells. In this work, using a highly sensitive fluorescence analysis, we have studied complexes of fluorescent analog of the natural heterocyclic antibiotic actinomycin D (7-aminoactinomycin D) with potential carriers: purine bases and fragmented DNA. The antibiotic is not only adsorbed on the surface ofpurine clusters, but also is embedded in them. The antibiotic is especially well integrated into the unwound DNA regions. Embedding is accompanied by a long-wavelength shift in the excitation spectrum. The magnitude of the shift was used for calculation of the interaction energy. In the case of AMD with guanine, caffeine and adenine, the value of energy was about of 7 kcal/mol and in the case of fragmented DNA it was only a bit higher: 7.7 kcal/mol. It can be assumed that guanine, adenine, caffeine, and the fragmented DNA could apply as carriers of antibiotic.
Topics: Anti-Bacterial Agents; DNA; DNA Fragmentation; Dactinomycin; Drug Delivery Systems; Humans; Neoplasms; Purines; Spectrometry, Fluorescence
PubMed: 25895354
DOI: 10.1134/s1068162014040074 -
Journal of Reproduction and Fertility Nov 1976Treatment of preimplantation mouse embryos in vitro with 10(-3) to 10(-1) mug actinomycin D/ml for 2 hr showed that (i) postimplantation development in vitro was...
Treatment of preimplantation mouse embryos in vitro with 10(-3) to 10(-1) mug actinomycin D/ml for 2 hr showed that (i) postimplantation development in vitro was inhibited most when embryos were treated at the morula stage and (ii) after the morula stage actinomycin D inhibited trophoblast outgrowth less than inner cell mass development.
Topics: Animals; Blastocyst; Dactinomycin; Embryo, Mammalian; Female; In Vitro Techniques; Mice; Superovulation; Trophoblasts
PubMed: 1033287
DOI: 10.1530/jrf.0.0480443 -
Cancer Jun 1973
Clinical Trial Randomized Controlled Trial
Topics: Adult; Aged; Dactinomycin; Female; Humans; Leukopenia; Male; Middle Aged; Remission, Spontaneous; Sarcoma, Kaposi; Thrombocytopenia; Vincristine
PubMed: 4709953
DOI: 10.1002/1097-0142(197306)31:6<1382::aid-cncr2820310613>3.0.co;2-d