• Title/Summary/Keyword: DNA binding

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Facilitated Protein-DNA Binding: Theory and Monte Carlo Simulation

  • Park, Ki-Hyun;Kim, Tae-Jun;Kim, Hyo-Joon
    • Bulletin of the Korean Chemical Society
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    • v.33 no.3
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    • pp.971-974
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    • 2012
  • The facilitated diffusion effect on protein-DNA binding is studied. A rigorous theoretical approach is presented to deal with the coupling between one-dimensional and three-dimensional diffusive motions. For a simplified model, the present approach can provide numerically exact results, which are confirmed by the lattice-based Monte Carlo simulations.

Novel TGACG-Motif Binding Protein of Soybean

  • Hong, Jong-Chan
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.40-47
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    • 1996
  • The promoters of a variety of plant genes are characterized by the presence of TGACG motif-containing sequences. These genes often exhibit quite diverse expression characteristics and in many case the TGACG-motif has been demonstrated to be essential for expression. Here we report the isolation and characterization of a soybean cDNA that encodes a novel basic/leucine zipper (bZIP) protein, STF1, that specifically interacts with Hex (TGACGTGG) and CRE (TGACGTCA) sequences. This protein contains a bZIP motif at C-teminus and an acidic domain at N-terminus. DNA binding specificities, heterodimer formation, and expression characteristics of STF1 were compared with a soybean TGA1 protein, STGA1. The soybean STF1 interacts with TGACG-sequences containing an ACGT core, while STGA1 requires TGACG as a sufficient binding sequence. The flanking sequences to the TGACG motif affected DNA binding of STF1 siginificantly. The STF1 mRNA is found mainly in dark grown soybean seedling with higher expression in apical and elongating hypocotyl, while STGA1 mRNA is highly abundant in roots of light grown plants. Furthermore, we demonstrate that STF1 heterodimerzes with G-box binding factorss (GBFs) which was not observed with TGA1. The fact that STF1 possesses both distinct DNA binding speficities and heterodimerization properties suggest that STF1 belongs to a new family of plant bZIP proteins which recognize the Hex/CRE motif.

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Single-strand DNA Binding of Actinomycin D with a Chromophore 2-Amino to 2-Hydroxyl Substitution

  • Yoo, Hoon;Rill, Randolph L.
    • BMB Reports
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    • v.36 no.3
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    • pp.305-311
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    • 2003
  • A modified actinomycin D was prepared with a hydroxyl group that replaced the amino group at the chromophore 2-position, a substitution known to strongly reduce affinity for double-stranded DNA. Interactions of the modified drug on single-stranded DNAs of the defined sequence were investigated. Competition assays showed that 2-hydroxyactinomycin D has low affinity for two oligonucleotides that have high affinities ($K_a\;=\;5-10{\times}10^6\;M^{-1}$ oligomer) for 7-aminoactinomycin D and actinomycin D. Primer extension inhibition assays performed on several single-stranded DNA templates totaling around 1000 nt in length detected a single high affinity site for 2-hydroxyactinomycin D, while many high affinity binding sites of unmodified actinomycin D were found on the same templates. The sequence selectivity of 2-hydroxyactinomycin D binding is unusually high and approximates the selectivity of restriction endonucleases. Binding appears to require a complex structure, including residues well removed from the polymerase pause site.

Antigenotoxicity of Ginseng Petroleum Ether Extract and its Action Mechanism (인삼 지용성성분인 유전독성억제효과와 작용기전)

  • 허문영
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.243-251
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    • 1998
  • Panax ginseng C.A. Meyer has been extensively used in the traditional oriental medicine as a restorative, tonic and prophylatic agent. Petroleum ether extract of panax ginseng C.A. Meyer (GPE) and its several fractions (PI-P5) were tested for the evaluation of antigenotoxicity against N-methyl-N-nitrosourea (MNU) and benzo(a)pyrene [B(a)P]-induced micronucleated reticulocytes in mouse peripheral blood. GPE and P2 showed more significant anticlastogenicity than other fractions did. To elucidate the anticlastogenic action mechanism of GPE and P2 against B(a)P, the alteration of B(a)P metabolism was studied. GPE and P2 inhibited B(a)P metabolism in the presence of 8-9 mix and decreased B(a)P-DNA binding in calf thymus DNA with 8-9 mix. They also decreased [$^3H$] MNU induced DNA binding and methylation to 7-methyl guanine and $O^{6}-methyl$ guanine adducts in calf thymus DNA by RPLC analysis. These results suggest that the anticlastogenicity of GPE and P2 on the B(a)P or MNU-induced clastogenicity is due to decrease of DNA binding with B(a)P or MNU, the inhibition of metabolism with B(a)P and the inhibition of methylation in DNA. Therefore, GPE and P2 may be useful chemopreventive agents of alkylating agent like MNU and secondary carcinogen like B(a)P.

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Physiological Function of a DNA-Binding Protein from Starved Cells in Combating Diverse External Stresses in Escherichia coli (대장균 세포 내 다양한 외부 스트레스에 대한 DPS 단백질의 생리적 기능)

  • Lee, Joo Hyeong;Cheong, Su Jin;Oh, Hun Taek;Kim, Woe Yeon;Jung, Young Jun
    • Journal of Life Science
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    • v.23 no.4
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    • pp.479-486
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    • 2013
  • The DNA-binding protein from starved cells (DPS), originally identified as a DNA binding protein in Escherichia coli, is known to play an important role in DNA protection. The aim of this study was to evaluate the functional roles of DPS in E. coli against various kinds of external stresses by comparing the properties of wild-type E. coli cells and dps knockout mutant E. coli (${\Delta}dps$) cells. Under various stress conditions, we measured the cell growth of the wild-type E. coli and the dps knockout mutant E. coli (${\Delta}dps$) cells using a UV spectrophotometer. The growth rate of the cells was compared to investigate the functional roles of the DPS protein in E. coli. In comparison to the properties of the wild-type E. coli cells, the dps knockout mutant E. coli (${\Delta}dps$) cells showed highly sensitive phenotypes under various stress conditions, such as heat shock, acidic pH, nutrient deficiency, and different concentrations of reactive oxygen species (ROS), suggesting that DPS plays key roles in E. coli in combating diverse external stresses. The DPS DNA-binding protein in E. coli plays crucial roles in bacterial cell growth and in the protection of the cells from environmental stresses by tightly binding and preserving their DNA molecules.

A Study of the Anticoagulatory DNA from the Earthworm, Lumbricus rubellus, and its Regulatory DNA-Binding Protein

  • Kim, Gyoung-Mi;Yu, Kyoung-Hee;Woo, Jeong-Im;Bahk, Yun-Kyoung;Paik, Seung R.;Kim, Jung-Gyu;Chang, Chung-Soon
    • BMB Reports
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    • v.32 no.6
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    • pp.567-572
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    • 1999
  • We have previously shown that a DNA fragment is responsible for the anticoagulatory effect of an earthworm, Lumbricus rubellus. The anticoagluant increased the activated partial thromboplastin time (APTT) and also inhibited the thrombin activity observed with either N-${\alpha}$-p-tosyl-L-arginine methyl ester (TAME) or H-D-phenyl-alanyl-L-pipecoil-L-arginine-p-nitroanilide (S-2238). Since trypsin digestion of the anticoagulant further increased the APTT, the possible presence of a regulatory protein for the anticoagulatory DNA was investigated by digesting the anticoagulant with trypsin and isolating the DNA fragment with C4-reversed phase HPLC. The DNA fragment lacking a regulatory protein was eluted in the flow-through fraction, and analyzed with thrombin and activated factor X. Activated factor X activity was more strongly inhibited than thrombin activity. For DNA digestion, we treated the anticoagulant with DNase and purified the DNA-binding protein with a FPLC Resource-S cation exchange column. The regulatory protein, with an $M_r$ of 55.0 kDa, reduced the anticoagulatory effect of the DNA fragment.

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Simple Screening Method for Double-strand DNA Binders Using Hairpin DNA-modified Magnetic Beads

  • Jo, Hun-Ho;Min, Kyoung-In;Song, Kyung-Mi;Ku, Ja-Kang;Han, Min-Su;Ban, Chang-Ill
    • Bulletin of the Korean Chemical Society
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    • v.32 no.1
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    • pp.247-250
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    • 2011
  • We designed an effective screening method for double strand DNA (dsDNA) binders using DNA-modified magnetic particles. Hairpin DNA was immobilized on the surface of magnetic particle for a simple screening of dsDNA binding materials in a solution containing various compounds. Through several magnetic separation and incubation processes, four DNA-binding materials, DAPI, 9AA, AQ2A, and DNR, were successfully screened from among five candidates. Efficiency of screening was demonstrated by HPLC analysis using a C2/18 reverse-phase column. In addition, their relative binding strengths were verified by measuring the melting temperature ($T_m$). If hairpin DNA sequence is modified for other uses, this magnetic bead-based approach can be applied as a high-throughput screening method for various functional materials such as anti-cancer drugs.

Inhibition of N-methyl-N-nitrosourea Induced Sister Chromatid Exchange and DNA Methylation by Galangin (N-Methyl-N-Nitrosourea 유도 자매염색분체교환생성과 DNA메칠화에 대한 Galangin의 억제효과)

  • 손수정;김정한;김영진;허인회;허문영
    • YAKHAK HOEJI
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    • v.39 no.1
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    • pp.94-101
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    • 1995
  • In order to evaluate the suppressive effects of galangin on the DNA damage induced by N-methyl-N-nitrosourea(MNU), in vitro sister chromatid exchange(SCE) test using Chinese Hamster ovary(CHO) cells was performed. Also the determinations of [$^{3}$H] MNU-induced total DNA binding and methylated DNA were performed to find out the mechanism of action by galangin. MNU-induced SCEs were significantly decreased by simultaneous and pretreatment of galangin when S-9 mix was added only. In post-treatment, however, the MNU-induced SCEs were not decreased when S-9 mix was added or not. [$^{3}$H] MNU-induced total DNA binding was significantly inhibited by the treatment of galangin in calf thymus DNA and CHO cells. HPLC analysis of DNA hydrolysates shows that galangin caused a dose-dependant decrease in calf thymus DNA, but not significant decrease in CHO cells. These results suggest that the inhibition of galangin on the MNU-induced SCEs is due to the decrease of DNA binding and methylation with MNU. Therefore, galangin may be useful as a chemopreventive agent of alkylating agents.

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Binding Modes of New Bis-Ru(II) Complexes to DNA: Effect of the Length of the Linker

  • Kwon, Byung-Hyang;Choi, Byung-Hoon;Lee, Hyun-Mee;Jang, Yoon-Jung;Lee, Jae-Cheol;Kim, Seog-K.
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1615-1620
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    • 2010
  • Bis-[dipyrido[3,2-$\alpha$:2',3'-c]phenazine)$_2$(1,10-phenanthroline)$_2Ru_2$]$^{2+}$ complexes (bis-Ru(II) complexes) tethered by linkers of various lengths were synthesized and their binding properties to DNA investigated by normal absorption and linear dichroism spectra, and fluorescence techniques in this study. Upon binding to DNA, the bis-Ru(II) complex with the longest linker (1,3-bis-(4-pyridyl)-propane), exhibited a negative $LD^r$ signal whose intensity was as large as that in the DNA absorption region, followed by a complicate $LD^r$ signal in the metal-to-ligand charge transfer region. The luminescence intensity of this bis-Ru(II) complex was enhanced. The observed $LD^r$ and luminescence results resembled that of the [Ru(1,10-phenanthroline)$_2$ dipyrido[3,2-$\alpha$:2',3'-c]phenazine]$^{2+}$ complex, whose dipyrido[3,2-$\alpha$:2',3'-c]phenazine (dppz) ligand has been known to intercalate between DNA bases. Hence, it is conclusive that both dppz ligands of the bis-Ru(II) complex intercalate. The binding stoichiometry, however, was a single intercalated dppz per ~ 2.3 bases, which violates the "nearest binding site exclusion" model for intercalation. The length between the two Ru(II) complexes may be barely long enough to accommodate one DNA base between the two dppz ligands, but not for two DNA bases. When the linker was shorter (4,4'-bipyridine or 1,2-bis-(4-pyridyl)-ethane), the magnitude of the LD in the dppz absorption region, as well as the luminescence intensity of both bis-Ru(II) complexes, was half that of the bis-Ru(II) complex bearing a long linker. This observation can be elucidated by a model whereby one of the dppz ligands intercalates while the other is exposed to the aqueous environment.