• Title/Summary/Keyword: DNA bending

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Single-Molecule Methods for Investigating the Double-Stranded DNA Bendability

  • Yeou, Sanghun;Lee, Nam Ki
    • Molecules and Cells
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    • v.45 no.1
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    • pp.33-40
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    • 2022
  • The various DNA-protein interactions associated with the expression of genetic information involve double-stranded DNA (dsDNA) bending. Due to the importance of the formation of the dsDNA bending structure, dsDNA bending properties have long been investigated in the biophysics field. Conventionally, DNA bendability is characterized by innate averaging data from bulk experiments. The advent of single-molecule methods, such as atomic force microscopy, optical and magnetic tweezers, tethered particle motion, and single-molecule fluorescence resonance energy transfer measurement, has provided valuable tools to investigate not only the static structures but also the dynamic properties of bent dsDNA. Here, we reviewed the single-molecule methods that have been used for investigating dsDNA bendability and new findings related to dsDNA bending. Single-molecule approaches are promising tools for revealing the unknown properties of dsDNA related to its bending, particularly in cells.

Mutant cAMP Receptor Protein Binds to DNA without DNA Bending (DNA 벤딩(휨) 없이 돌연변이 cAMP 수용체 단백질의 결합)

  • Gang, Jong-Back
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1225-1228
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    • 2006
  • Cyclic AMP receptor protein (CRP) complexed with cAMP binds to DNA and induces sharp DNA bending around ${\sim}90$ degree. Previous publication (5), however, reported that mutant CRP:cGMP complex showed high migration rate relative to mutant CRP:cAMP complex on native polyacrylamide gel. To confirm DNA structural change in the presence of CRP and cyclic nucleotide, molar cyclization factor $(j_M)$ [13] was measured with 6 constructed DNA fragments. Nonlinear regression analysis of $j_M$ data indicated that mutant CRP did not induce DNA bending in the presence of cGMP but bent DNA in the presence of cAMP without any helical twist change in DNA.

Bending of DNA by cAMP Receptor Protein(CRP)

  • 정수열
    • Journal of Life Science
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    • v.4 no.3
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    • pp.119-123
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    • 1994
  • CRP에 의한 DNA bending의 첫 실험은 cAMP, CRP, DNA complex의 gelelectrophoresis에 의해 확인되었다. Bent DNA fragment의 이동은 같은 크기의 linear fragment보다 느리며, EH cAMP, CRP, DNA 결합위치에 따라서 상이한 이동 pattern을 나타낸다. 즉, DNA 단편의 중앙에 CRP binding site가 있을 때는 말단부분에 있을 때보다 이동도가 느리다. benting각의 크기는 일반적으로 안정한 복합체의 형성에 관계하며, 큰 각을 갖는 것이 보다 더 안정된 구조를 형성하여 전사가 촉진되는 것으로 밝혀져 있다. 본고에서는 CRP에 의한 bending과 CRP에 의한 전사조절 관계와 bending의 관계에 대하여 알아보았다.

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Helical Periodicity of $(dT)_n{\cdot}(dA)_n{\cdot}(dT)_n$ Triple - Stranded DNA

  • Kim, Ki-Hyun;Koo, Hyeon-Sook
    • BMB Reports
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    • v.30 no.6
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    • pp.426-430
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    • 1997
  • The helical periodicity of the triple-stranded $(dT)_n{\cdot}(dA)_n{\cdot}(dT)_n$ sequence was determined by measuring gel-mobilities of bent DNA fragments containing the sequence. In the bent DNA fragments, a $GA_{22}G$ $CT_{22}C$ sequence was located between two bent DNA loci composed of six $A_{6}{\cdot}T_{6}$ repeats. and the DNA length between the bent DNA loci was varied by 1 base pair over a full helical turn. The gel mobility of each bent DNA fragment reflected the overall extent of DNA bending and varied with the DNA length between the two bent loci. Mobilities of the bent DNA fragments in 5% polyacrylamide gel were measured after preincubating the DNA fragments both in the presence and absence of $CT_{22}C$ oligonucleotide. By comparing the bent DNA fragments containing an intermolecular triplex structure with those of a genuine duplex structure in the gel mobilities, the helical periodicity of the $T_n{\cdot}A_n{\cdot}T_n$ triplex DNA was determined to be $11.5({\pm}0.3)bp/turn$.

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RBF-POD reduced-order modeling of DNA molecules under stretching and bending

  • Lee, Chung-Hao;Chen, Jiun-Shyan
    • Interaction and multiscale mechanics
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    • v.6 no.4
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    • pp.395-409
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    • 2013
  • Molecular dynamics (MD) systems are highly nonlinear and nonlocal, and the conventional model order reduction methods are ineffective for MD systems. The RBF-POD method (Lee and Chen, 2013) employed a radial basis function (RBF) approximated potential energies and inter-atomic forces of MD systems under the framework of the proper orthogonal decomposition (POD) method for the reduced-order modeling of MD systems. In this work, we focus on the numerical procedures of the RBF-POD method and demonstrate how to apply this approach to the modeling of ds-DNA molecules under stretching and bending conditions.

DNA Binding Specificity of Proteus mirabilis Transcription Regulator (Proteus mirabilis 전사 조절 단백질의 DNA 결합 특성)

  • Gang, Jong-Back
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.158-162
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    • 2011
  • Amino acid sequence alignment shows that $\underline{P}$roteus $\underline{m}$irabilis $\underline{t}$ranscription $\underline{r}$egulator (PMTR) has cystein sequence homology at metal binding domain to CueR (copper resistance) protein, which conserves two cysteins (Cys 112 and Cys 120 in PMTR). Gel shift assay revealed that PMTR protein bound to promoter region of Escherichia coli copA (copper-translocating P-type ATPase) and Proteus mirabilis atpase (putative copper-translocating P-type ATPase) genes except that of E. coli zntA (zinc-translocating P-type ATPase) gene. DNase I protection experiment indicated that PMTR protein protected the region over -35 box and close to -10 box. DNase I hypersensitive bases were shown at C and A bases of labeled template strand and at G and C bases of labeled non-template strand of DNA. These hypersensitive bases were appeared in other metalloregulatory proteins of MerR family, which suggests protein-induced DNA bending.

Ionic Strength Dependent Binding Mode of 9-Aminoacridine to DNA

  • 김혜경;조태섭;Kim, Seog K.
    • Bulletin of the Korean Chemical Society
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    • v.17 no.4
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    • pp.358-362
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    • 1996
  • The ionic strength dependent binding mode of 9-aminoacridine (9AA), a well-known DNA intercalator, to DNA is studied by flow linear dichroism, circular dichroism, fluorescence techniques and equilibrium dialysis. The DNA-bound 9AA exhibits spectral properties corresponding to the intercalative binding mode disregarding the salt concentrations; the angle between the long-axis transition moment of the 9AA molecule and DNA helix axis is calculated to be about 65°, indicating a significant deviation from the classical intercalation. At low salt concentrations, however, upwards bending curve in Stern-Volmer plot is observed (where 9AA is a fluorophore and DNA a quencher), indicating the coexistence of both static and dynamic quenching mechanisms or the existence of an additional binding site.

Study on the chemical environment for conformational change of i-motif DNA by atomic force microscopy cantilever (AFM 캔틸레버를 이용한 i-motif DNA의 구조 변화에 미치는 화학적 환경에 대한 연구)

  • Jung, Hwi-Hun;Park, Jin-Sung;Yang, Jae-Moon;Lee, Sang-Woo;Eom, Kil-Ho;Kwon, Tae-Yun;Yoon, Dae-Sung
    • Journal of Sensor Science and Technology
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    • v.19 no.3
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    • pp.214-220
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    • 2010
  • Three-dimensional(3D) structure of specific DNA can be changed between two conformations under an external environmental transition such as pH and salt concentration variations. We have experimentally observed the conformational transitions of i-motif DNA using AFM cantilever bioassay. It is shown that pH change of a solvent induces the bending defleciton change of a cantilever functionalized by i-motif DNA. This indicates that cantilever bioassay enables the label-free detection of DNA structural changes upon pH change. It is implied that cantilever bioassay can be a de novo route to quantitatively understand the conformational transitions of biological molecules under environmental changes.

Binding Aspect of Cyclic AMP Receptor Protein to Symmetrically Synthetic 22-, 28- and 30-Base-Pair lac Promoters

  • Park, Sang-Ho;Lee, Tae-Woo;Hwang, Eun-Suk;Lee, Seung-Ki;Shin, Cha-Gyun;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.1 no.1
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    • pp.31-44
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    • 1997
  • The effect of the binding of CRP to the symmetrically synthetic 22, 28, and 30 bp lac promoter was investigated by 1H NMR. The binding of cAMP*CRP to the 22 bp DNA did not bring about any changes in the chemical shift values, but did cause selective line broadening of imino proton resonances of specific base pairs. However, The binding of cAMP*CRP to the 28 and 30 bp DNA brought about large changes on the imino proton resonances that seems to be induced by DNA bending. We studied also the role of cAMP as an activator of DNA/CRP complex formation by gel mobility shift assay. Gel mobility shift assay revealed that the cAMP*CRP complex was not able to bind to the 22 bp DNA fragment, but was able to bind to the 28 bp DNA fragment of lac promoter region.

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Tethered DNA shear dynamics in the flow gradient plane: application to double tethering

  • Lueth, Christopher A.;Shaqfeh, Eric S.G.
    • Korea-Australia Rheology Journal
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    • v.19 no.3
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    • pp.141-146
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    • 2007
  • We examine the wall contact of a $3\;{\mu}m$ tethered DNA chain's free end under shear with a focus on developing schemes for double-tethering in the application of making scaffolds for molecular wires. At this scale our results are found to be highly dependent on small length scale rigidity. Chain-end-wall contact frequency, mean fractional extension deficit upon contact, and standard deviation in extension upon contact are examined for scaling with dimensionless flow strength, Wi. Predictions made using a one dimensional approximation to the Smoluchowski equation for a dumbbell and three dimensional dumbbell simulations produce extension deficit, standard deviation, and frequency scaling exponents of -1/3, -1/3, and 2/3, respectively whereas more fine-grained Kratky-Porod (KP) simulations produce scaling exponents of -0.48, -0.42, and 0.76. The contact frequency scaling of 2/3 is derived from the known results regarding cyclic dynamics Analytical scaling predictions are in agreement with those previously proposed for ${\lambda}-DNA$. [Ladoux and Doyle, 2000, Doyle et al., 2000]. Our results suggest that the differences between the dumbbell and the KP model are associated with the addition of chain discretization and the correct bending potential in the latter. These scaling results will aide future exploration in double tethering of DNA to a surface.