• 제목/요약/키워드: DNA interaction

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Paramyrothecium eichhorniae sp. nov., Causing Leaf Blight Disease of Water Hyacinth from Thailand

  • Pinruan, Umpawa;Unartngam, Jintana;Unartngam, Arm;Piyaboon, Orawan;Sommai, Sujinda;Khamsuntorn, Phongsawat
    • Mycobiology
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    • 제50권1호
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    • pp.12-19
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    • 2022
  • Paramyrothecium eichhorniae sp. nov. was observed and collected from Chiang Mai and Phetchaburi Provinces, Thailand. This new species is introduced based on morphological and molecular evidence. This fungus is characterized by its production of sporodochium conidiomata with a white setose fringe surrounding an olivaceous green to dark green slimy mass of conidia, penicillately branched conidiophores, and aseptate and cylindrical to ellipsoid conidia. Phylogenetic analyses of combined LSU rDNA, ITS rDNA, tef1, rpb2, tub2 and cmdA sequence data using maximum parsimony, maximum likelihood and Bayesian approaches placed the fungus in a strongly supported clade with other Paramyrothecium species in Stachybotryaceae (Hypocreales, Sordariomycetes). The descriptions of the species are accompanied by illustrations of morphological features, and a discussion of the related taxa is presented.

사람 NC-37 세포에서 Benzo(a)pyrene과 Doxorubicin에 의한 Nuclei내전과 c-myc 유전자의 발현에 대한 연구 (Studies on the Nuclei Adduction and Expression of c-myc Gene by Benzo(a)pyrene and Doxorubicin in Human NC-37 Cells)

  • 김호찬;정인철;조무연
    • 생명과학회지
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    • 제8권4호
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    • pp.400-409
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    • 1998
  • Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

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Label/Quencher-Free Detection of Exon Deletion Mutation in Epidermal Growth Factor Receptor Gene Using G-Quadruplex-Inducing DNA Probe

  • Kim, Hyo Ryoung;Lee, Il Joon;Kim, Dong-Eun
    • Journal of Microbiology and Biotechnology
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    • 제27권1호
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    • pp.72-76
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    • 2017
  • Detection of exon 19 deletion mutation in the epidermal growth factor receptor (EGFR) gene, which results in increased and sustained phosphorylation of EGFR, is important for diagnosis and treatment guidelines in non-small-cell lung cancer. Here, we have developed a simple and convenient detection system using the interaction between G-quadruplex and fluorophore thioflavin T (ThT) for discriminating EGFR exon 19 deletion mutant DNA from wild type without a label and quencher. In the presence of exon 19 deletion mutant DNA, the probe DNAs annealed to the target sequences were transformed into G-quadruplex structure. Subsequent intercalation of ThT into the G-quadruplex resulted in a light-up fluorescence signal, which reflects the amount of mutant DNA. Due to stark differences in fluorescence intensity between mutant and wild-type DNA, we suggest that the induced G-quadruplex structure in the probe DNA can report the presence of cancer-causing deletion mutant DNAs with high sensitivity.

The stimulatory effect of CaCl2, NaCl and NH4NO3 salts on the ssDNA-binding activity of RecA depends on nucleotide cofactor and buffer pH

  • Ziemienowicz, Alicja;Rahavi, Seyed Mohammad Reza;Kovalchuk, Igor
    • BMB Reports
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    • 제44권5호
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    • pp.341-346
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    • 2011
  • The single-stranded DNA binding activity of the Escherichia coli RecA protein is crucial for homologous recombination to occur. This and other biochemical activities of ssDNA binding proteins may be affected by various factors. In this study, we analyzed the effect of $CaCl_2$, NaCl and $NH_4NO_3$ salts in combination with the pH and nucleotide cofactor effect on the ssDNA-binding activity of RecA. The studies revealed that, in addition to the inhibitory effect, these salts exert also a stimulatory effect on RecA. These effects occur only under very strict conditions, and the presence or absence and the type of nucleotide cofactor play here a major role. It was observed that in contrast to ATP, ATP${\gamma}$S prevented the inhibitory effect of NaCl and $NH_4NO_3$, even at very high salt concentration. These results indicate that ATP${\gamma}$S most likely stabilizes the structure of RecA required for DNA binding, making it resistant to high salt concentrations.

Direction of Intercalation of a bis-Ru(II) Complex to DNA Probed by a Minor Groove Binding Molecule 4',6-Diamidino-2-phenylindole

  • Jang, Yoon Jung;Kim, Raeyeong;Chitrapriya, Nataraj;Han, Sung Wook;Kim, Seog K.;Bae, Inho
    • Bulletin of the Korean Chemical Society
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    • 제34권10호
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    • pp.2895-2899
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    • 2013
  • Direction of intercalation to DNA of the planar dipyrido[3,2-a:2',3'-c]phenazine ligands (dppz) of a bis-Ru(II) complex namely, $[Ru(1,10-phenanthroline)_2dipyrido[3,2-a:2^{\prime},3^{\prime}-c]phenazine]^{2+}$ linkered by a 1,3-bis(4-pyridyl)propane, was investigated by probing the behavior of 4',6-diamidino-2-phenylindole (DAPI) that bound deep in the minor groove. Bis-intercalation of DPPZ resulted in a little blue shift and hyperchromism in DAPI absorption band, and a large decrease in DAPI fluorescence intensity which accompined by an increase in the dppz emission intensity. Diminishing the intenisty of the positive induced circular dichroism (CD) and linear dichroism (LD) were also observed. These spectral changes indicated that insertion of dppz ligand caused the change of the binding mode of DAPI, which probably moved to the exterior of DNA from the minor groove and interacted with the phospghate groups of DNA by electrostatic interaction. At the surface of DNA, DAPI binds at the phosphate groups of DNA by electrostatic attraction. Consequently, this observation indicated that the dppz ligand intercalated from the minor groove.

DNA-Binding and Thermodynamic Parameters, Structure and Cytotoxicity of Newly Designed Platinum(II) and Palladium(II) Anti-Tumor Complexes

  • Mansouri-Torshizi, Hassan;Saeidifar, Maryam;Khosravi, Fatemeh;Divsalar, Adeleh;Saboury, Ali.Akbar;Ghasemi, Zahra Yekke
    • Bulletin of the Korean Chemical Society
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    • 제32권3호
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    • pp.947-955
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    • 2011
  • The complexes [Pd(bpy)(Hex-dtc)]$NO_3$ and [Pt(bpy)(Hex-dtc)]$NO_3$ (bpy is 2,2'-bipyridine and Hex-dtc is hexyldithiocarbamato ligands) were synthesized and characterized by elemental analysis and spectroscopic studies. The cytotoxicity assay of the complexes has been performed on chronic myelogenous leukemia cell line, K562, at micromolar concentration. Both complexes showed cytotoxic activity far better than that of cisplatin under the same experimental conditions. The binding parameters of the complexes with calf thymus DNA (CT-DNA) was investigated using UV-visible and fluorescence techniques. They show the ability of cooperatively intercalating in CT-DNA. Gel filtration studies demonstrated that platinum complex could cleave the DNA. In the interaction studies between the Pd(II) and Pt(II) complexes with CT-DNA, several binding and thermodynamic parameters have been determined, which may provide deeper insights into the mechanism of action of these types of complexes with nucleic acids.

R3V6 Amphiphilic Peptide with High Mobility Group Box 1A Domain as an Efficient Carrier for Gene Delivery

  • Ryu, Jaehwan;Jeon, Pureum;Lee, Minhyung
    • Bulletin of the Korean Chemical Society
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    • 제34권12호
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    • pp.3665-3670
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    • 2013
  • The R3V6 peptide includes a hydrophilic arginine stretch and a hydrophobic valine stretch. In previous studies, the R3V6 peptide was evaluated as a gene carrier and was found to have low cytotoxicity. However, the transfection efficiency of R3V6 was lower than that of poly-L-lysine (PLL) in N2A neuroblastoma cells. In this study, the transfection efficiency of R3V6 was improved in combination with high mobility group box 1A domain (HMGA). HMGA is originated from the nuclear protein and has many positively-charged amino acids. Therefore, HMGA binds to DNA via charge interaction. In addition, HMGA has a nuclear localization signal peptide and may increase the delivery efficiency of DNA into the nucleus. The ternary complex with HMGA, R3V6, and DNA was prepared and evaluated as a gene carrier. First, the HMGA/DNA complex was prepared with a negative surface charge. Then, R3V6 was added to the complex to coat the negative charges of the HMGA/DNA complex, forming the ternary complex of HMGA, R3V6, and DNA. A physical characterization study showed that the ternary complex was more stable than the PLL/DNA complex. The HMGA/R3V6/DNA complex had a higher transfection efficiency than the PLL/DNA, HMGA/DNA, or R3V6/DNA complexes in N2A cells. Furthermore, the HMGA/R3V6/DNA complex was not toxic to cells. Therefore, the HMGA/R3V6/DNA complex may be a useful gene delivery carrier.

In silico docking of methyl isocyanate (MIC) and its hydrolytic product (1, 3-dimethylurea) shows significant interaction with DNA Methyltransferase 1 suggests cancer risk in Bhopal-Gas-Tragedy survivors

  • Khan, Inbesat;Senthilkumar, Chinnu Sugavanam;Upadhyay, Nisha;Singh, Hemant;Sachdeva, Meenu;Jatawa, Suresh Kumar;Tiwari, Archana
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권17호
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    • pp.7663-7670
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    • 2015
  • DNA methyltransferase 1 (DNMT1) is a relatively large protein family responsible for maintenance of normal methylation, cell growth and survival in mammals. Toxic industrial chemical exposure associated methylation misregulation has been shown to have epigenetic influence. Such misregulation could effectively contribute to cancer development and progression. Methyl isocyanate (MIC) is a noxious industrial chemical used extensively in the production of carbamate pesticides. We here applied an in silico molecular docking approach to study the interaction of MIC with diverse domains of DNMT1, to predict cancer risk in the Bhopal population exposed to MIC during 1984. For the first time, we investigated the interaction of MIC and its hydrolytic product (1,3-dimethylurea) with DNMT1 interacting (such as DMAP1, RFTS, and CXXC) and catalytic (SAM, SAH, and Sinefungin) domains using computer simulations. The results of the present study showed a potential interaction of MIC and 1,3-dimethylurea with these domains. Obviously, strong binding of MIC with DNMT1 interrupting normal methylation will lead to epigenetic alterations in the exposed humans. We suggest therefore that the MIC-exposed individuals surviving after 1984 disaster have excess risk of cancer, which can be attributed to alterations in their epigenome. Our findings will help in better understanding the underlying epigenetic mechanisms in humans exposed to MIC.

EFFECTS OF LYSINE AND ENERGY LEVELS ON GROWTH PERFORMANCE THIGH MUSCLE COMPOSITION AND UTILIZATION OF NUTRIENTS IN BROILER CHICKS

  • Park, B.C.;Han, I.K.;Choi, Y.J.;Yun, C.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제5권1호
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    • pp.129-138
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    • 1992
  • The effects of dietary levels of lysine and energy on growth performance, the content of DNA, RNA and protein in liver, thigh muscle composition and nutrient utilization in broiler chicks were investigated in an experiment involvies with 2 levels of dietary energy : 3,200 (2900) 2,900 (2700) kcal ME/kg) and 6 levels of lysine : 0.6(0.5), 0.8(0.7), 1.0(0.9), 1.2(1.1), 1.4(1.3), and 1.6(1.5)% was carried out. A total number of 384 male broiler chicks was used for a period of 7 weeks. Body weight gain of 1.0(0.9)% lysine level group was significantly (p < 0.01) higher than that of any other groups. Interaction between lysine and energy in the feed intake was observed (p < 0.05). Present data indicate that the content of DNA in liver tissues was significantly (p < 0.05) different by the levels of lysine, namely, 1.0(0.9)% or 1.2(1.1)% lysine level groups showed higher content than other groups (p < 0.01). Dietary levels of 1.2(1.1)% or 1.6(1.5)% lysine groups showed the highest protein content in thigh muscle tissues than that of any other groups (p < 0.05). Interaction between energy and lysine in the content of protein of thigh muscle tissues was shown (p < 0.01). The level of 0.6% lysine group showed the highest fat content in thigh muscle tissues than any other groups. Interaction between lysine and energy in the content of crude ash and crude fat of thigh muscle tissues was observed (p < 0.01). Apparent amino acid availability of arginine, glycine and threonine (p < 0.01), phenylalanine (p < 0.05) were significantly affected by the levels of lysine and interaction between lysine and energy was found only in arginine (p < 0.01).

Effects of the Heptasequence SPTSPTY of Rat Nuclear Factor 1-A on Interactions between the C-Terminal Regions of Mammalian Nuclear Factor 1 Proteins

  • Hwang, Jung-Su;Kim, Ji-Young
    • BMB Reports
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    • 제33권6호
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    • pp.519-524
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    • 2000
  • NF1 proteins are a family of DNA binding proteins which consist of two separate domains, N-terminal DNA binding domain and C-terminal transcription activation domain. The N-terminal 220 amino acids are highly conserved and are also known to mediate dimerization of NF1 proteins. The C-terminal regions of different type of NF1 proteins are heterogeneous and responsible for transcriptional activation. In this study, we tested the interaction between different domains of rat NF1-A protein by yeast two hybrid analysis and observed the interaction between C-terminal regions of NF1-A which do not contain the N-terminal dimerization domain. Our results showed that the C-terminal region of rat NF1-A between residues 231 and 509 strongly interacted not only with itself, but also with human NF1/CTF1 which is a different type of NF1. When the C-terminal region was divided into two fragments, one from residue 231 to 447 and the other from 448 to 509, the two fragments were able to interact with the C-terminal region of NF1-A significantly. This indicates that both fragments contain independent interaction domains. Analysis of the interactions with alanine substituted fragments showed that substitutions of the heptasequence, SPTSPTY of NF1-A, affected interaction between NF1 proteins. Our results strongly suggest that C-terminal regions may also be important for the formation of homo- and heterodimers in addition to the N-terminal dimerization domain. Also, the heptasequence motif may play some roles in dimer formation.

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