• Title/Summary/Keyword: DNA molecules

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Crystal Structures of the Two Isomorphous A-DNA Decamers d(GTACGCGTAC) and d(GGCCGCGGCC)

  • Kim, Tae-gyun;Kwon, Taek-Hun;Jung, Hye-sun;Ku, Ja-Kang;Sundaralingam, Muttaiya;Ban, Chang-ill
    • Bulletin of the Korean Chemical Society
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    • v.27 no.4
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    • pp.568-572
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    • 2006
  • To study the effect of sequence on DNA structure, the two decamer crystal structures one alternating,d(GTACGCGTAC), and the other non-alternating, d(GGCCGCGGCC), were solved. Crystals of both decamers belong to the hexagonal space group $P6_122$, with one strand in the asymmetric unit. The unit cell constants of the alternating decamer are a = b = 39.26 $\AA$, c = 77.70 $\AA$. The structure was refined with 1,828 reflections from 8.0 to 2.0 Aresolution to an R value of 21.3% with all DNA atoms and 63 water molecules. The isomorphous non-alternating decamer had unit cell dimensions of a = b = 39.05 $\AA$, c = 82.15 $\AA$. The structure was refined with 2,423 reflections from 8.0 to 2.0 $\AA$ resolution to a final R value of 22.2% for all DNA atoms and 65 water molecules. Although the average helical parameters of the decamers are typical of A-DNAs, there are some minor differences between them. The helical twist, rise, x-displacement, inclination and roll alternate in the alternating decamer, but do not in the non-alternating decamer. The backbone conformations in both structures show some differences; the residue G(7) of the alternating decamer is trans for $\alpha$ and $\gamma$ while the trans conformations are observed at the residue G(8) of the non-alternating decamer.

Ordered Fragmentation of pDNA induced by PEG-PLL block copolymer -Correlation between Condensation degree and Biological Activity by Cell-Free System-

  • Osada, Kensuke;Doi, Motoyoshi;Shiotani, Tomonori;Yamasaki, Yuichi;Kataoka, Kazunori
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.254-254
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    • 2006
  • The sensitivity of plasmid DNA (pDNA) to S1 nuclease, an enzyme to cleave a single-strand DNA, was dramatically modulated through a supramolecular assembly (polyion complex micelle) with a synthetic block copolymer, poly(ethylene glycol)-b-poly(L-lysine) (PEG-PLL). The pDNA condensed in stoichiometric charge ratio was cleaved into 7 fragments each being 10/12, 9/12, 8/12, 6/12, 4/12, 3/12, and 2/12 of the original DNA length, on the other hand, the pDNA condensed in higher charge ratios (>4), were digested into non-specific manner. Condensation of the pDNA was investigated from two viewpoints that how does the rigid DNA molecules fold and condense and how does the condensation influence their biological activity.

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Solving the Monkey and Banana Problem Using DNA Computing (DNA 컴퓨팅을 이용한 원숭이와 바나나 문제 해결)

  • 박의준;이인희;장병탁
    • Korean Journal of Cognitive Science
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    • v.14 no.2
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    • pp.15-25
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    • 2003
  • The Monkey and Banana Problem is an example commonly used for illustrating simple problem solving. It can be solved by conventional approaches, but this requires a procedural aspect when inferences are processed, and this fact works as a limitation condition in solving complex problems. However, if we use DNA computing methods which are naturally able to realize massive parallel processing. the Monkey and Banana Problem can be solved effectively without weakening the fundamental aims above. In this paper, we design a method of representing the problem using DNA molecules, and show that various solutions are generated through computer-simulations based on the design. The simulation results are obviously interesting in that these are contrary to the fact that the Prolog program for the Monkey and Banana Problem, which was implemented from the conventional point of view, gives us only one optimal solution. That is, DNA computing overcomes the limitations of conventional approaches.

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Hepatitis B Virus X Protein Stimulates Virus Replication Via DNA Methylation of the C-1619 in Covalently Closed Circular DNA

  • Lee, Hyehyeon;Jeong, Hyerin;Lee, Sun Young;Kim, Soo Shin;Jang, Kyung Lib
    • Molecules and Cells
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    • v.42 no.1
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    • pp.67-78
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    • 2019
  • Methylation of HBV cccDNA has been detected in vivo and in vitro; however, the mechanism and its effects on HBV replication remain unclear. HBx derived from a 1.2-mer HBV replicon upregulated protein levels and enzyme activities of DNA methyltransferase 1 (DNMT1), 3a, and 3b, resulting in methylation of the negative regulatory region (NRE) in cccDNA, while none of these effects were observed with an HBx-null mutant. The HBx-positive HBV cccDNA expressed higher levels of HBc and produced about 4-fold higher levels of HBV particles than those from the HBx-null counterpart. For these effects, HBx interrupted the action of NRE binding protein via methylation of the C-1619 within NRE, resulting in activation of the core promoter. Treatment with 5-Aza-2′dC or DNMT1 knock-down drastically impaired the ability of HBx to activate the core promoter and stimulate HBV replication in 1.2-mer HBV replicon and in vitro infection systems, indicating the positive role of HBx-mediated cccDNA methylation in HBV replication.

Photocyclodimerization of Maleimide

  • Shim, Sang-Chul;Bong, Pill-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.3 no.3
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    • pp.115-119
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    • 1982
  • The photoreaction of maleimide, one of the best model compounds of DNA molecules for psoralen-DNA photoreactions, is studied in order to investigate the photoreactivity and the mechanism of the maleimide-psoralen photoreaction. The (2+2) photocyclodimer of maleimide was obtained in solution state by direct or sensitized irradiation. The rate constant of dimerization is determined by quenching studies and found to be of the order of $10^9 M^{-1}sec^{-1}$. The direct dimerization of maleimide is found to undergo through the triplet excited state. The quantum yields of dimerization are dependent on the maleimide concentration.

Enhancing Transfection Efficiency Using Polyethylene Glycol Grafted Polyethylenimine and Fusogenic Peptide

  • Lee, Haeshin;Jeong, Ji-Hoon;Lee, Je-Hoon;Park, Tae-Gwan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.269-273
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    • 2001
  • This study presents a new formulation method for improving DNA transfection effi-ciency using a fusogenic peptide and polyethylene glycol-grafted polyethylenimine. Succinimidyls succinate polyethylene glycol (PEG-SSA) was conjugated with polyethylenimine(PEL). PEL is well known for a good endosomal escaping and DNA condensign agent. The positively charged syn-thetic fusogenic peptide, KALA was coated on the negatively charged PEG-g-PEI/DNA and PEI/DNA complexes. The KALA/PEI/ DNA complexes exhibited aggregation behavior at higher KALA coating amount with an effective diameter of around 1,000 nm. However, the LALA/PEG-g-PEI/DNA complexes were 100-300 nm in size with a surface zeta-potential (ζ)value of about +20mV. The conjugated PEG molecules suppressed any KALA-mediated inter-particle aggregation, and thereby improved the transfection efficiency, Consequently, the transfection efficiency of the KALA/PEG-g-PEI/DNA complexes was obtained by utilizing both the fusogenic activity of KALA and the steric repulsion effect of PEC.

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Identification of Immunostimulatory Oligodeoxynucleotide from Escherichia coli Genomic DNA

  • Choi, Yong-Jun;Lee, Keun-Wook;Kwon, Hyung-Joo;Kim, Doo-Sik
    • BMB Reports
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    • v.39 no.6
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    • pp.788-793
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    • 2006
  • Bacterial DNA containing immunostimulatory CpG motifs can stimulate antigen-presenting cells to express co-stimulatory molecules and to produce various cytokines in vivo and in vitro. In this study, we fragmented macromolecular E.coli genomic DNA with DNase I, and analyzed the ability of the resulting DNA fragments to induce the NF-${\kappa}B$ activation and humoral immune response. Furthermore, using computational analysis and luciferase assay for synthetic ODNs based on the sequence of the immunostimulatory DNA fragments (DF-ODNs), an active component of DF-ODNs sequences was investigated. Experimental results demonstrated that DF-ODN is optimal for the NF-${\kappa}B$-responsive promoter activation in the mouse macrophage cell line and the humoral immune response in vivo. In agreement with the activity of the DF-ODNs processed by DNase I, a synthetic ODN based on the DF-ODN sequences is potent at inducing IL-12 mRNA expression in primary dendritic cells. These results suggest that the discovery and characterization of a highly active natural CpG-ODN may be achieved by the analyses of bacterial DNA fragments generated by a nuclease activity.

Cellular Dynamics of Rad51 and Rad54 in Response to Postreplicative Stress and DNA Damage in HeLa Cells

  • Choi, Eui-Hwan;Yoon, Seobin;Hahn, Yoonsoo;Kim, Keun P.
    • Molecules and Cells
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    • v.40 no.2
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    • pp.143-150
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    • 2017
  • Homologous recombination (HR) is necessary for maintenance of genomic integrity and prevention of various mutations in tumor suppressor genes and proto-oncogenes. Rad51 and Rad54 are key HR factors that cope with replication stress and DNA breaks in eukaryotes. Rad51 binds to single-stranded DNA (ssDNA) to form the presynaptic filament that promotes a homology search and DNA strand exchange, and Rad54 stimulates the strand-pairing function of Rad51. Here, we studied the molecular dynamics of Rad51 and Rad54 during the cell cycle of HeLa cells. These cells constitutively express Rad51 and Rad54 throughout the entire cell cycle, and the formation of foci immediately increased in response to various types of DNA damage and replication stress, except for caffeine, which suppressed the Rad51-dependent HR pathway. Depletion of Rad51 caused severe defects in response to postreplicative stress. Accordingly, HeLa cells were arrested at the G2-M transition although a small amount of Rad51 was steadily maintained in HeLa cells. Our results suggest that cell cycle progression and proliferation of HeLa cells can be tightly controlled by the abundance of HR proteins, which are essential for the rapid response to postreplicative stress and DNA damage stress.

Identification of Protein Phosphatase 4 Inhibitory Protein That Plays an Indispensable Role in DNA Damage Response

  • Park, Jaehong;Lee, Jihye;Lee, Dong-Hyun
    • Molecules and Cells
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    • v.42 no.7
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    • pp.546-556
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    • 2019
  • Protein phosphatase 4 (PP4) is a crucial protein complex that plays an important role in DNA damage response (DDR), including DNA repair, cell cycle arrest and apoptosis. Despite the significance of PP4, the mechanism by which PP4 is regulated remains to be elucidated. Here, we identified a novel PP4 inhibitor, protein phosphatase 4 inhibitory protein (PP4IP) and elucidated its cellular functions. PP4IP-knockout cells were generated using the CRISPR/Cas9 system, and the phosphorylation status of PP4 substrates (H2AX, KAP1, and RPA2) was analyzed. Then we investigated that how PP4IP affects the cellular functions of PP4 by immunoprecipitation, immunofluorescence, and DNA double-strand break (DSB) repair assays. PP4IP interacts with PP4 complex, which is affected by DNA damage and cell cycle progression and decreases the dephosphorylational activity of PP4. Both overexpression and depletion of PP4IP impairs DSB repairs and sensitizes cells to genotoxic stress, suggesting timely inhibition of PP4 to be indispensable for cells in responding to DNA damage. Our results identify a novel inhibitor of PP4 that inhibits PP4-mediated cellular functions and establish the physiological importance of this regulation. In addition, PP4IP might be developed as potential therapeutic reagents for targeting tumors particularly with high level of PP4C expression.

Code Optimization in DNA Computing for the Hamiltonian Path Problem (해밀톤 경로 문제를 위한 DNA 컴퓨팅에서 코드 최적화)

  • 김은경;이상용
    • Journal of KIISE:Software and Applications
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    • v.31 no.4
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    • pp.387-393
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    • 2004
  • DNA computing is technology that applies immense parallel castle of living body molecules into information processing technology, and has used to solve NP-complete problems. However, there are problems which do not look for solutions and take much time when only DNA computing technology solves NP-complete problems. In this paper we proposed an algorithm called ACO(Algorithm for Code Optimization) that can efficiently express DNA sequence and create good codes through composition and separation processes as many as the numbers of reaction by DNA coding method. Also, we applied ACO to Hamiltonian path problem of NP-complete problems. As a result, ACO could express DNA codes of variable lengths more efficiently than Adleman's DNA computing algorithm could. In addition, compared to Adleman's DNA computing algorithm, ACO could reduce search time and biological error rate by 50% and could search for accurate paths in a short time.