• Title/Summary/Keyword: DNA mobility

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The Regulation of AP-1 DNA Binding Activity by Long-term Nicotine Stimulation in Bovine Adrenal Medullary Chromaffin Cells: Role of Second Messengers

  • Lee, Jin-Koo;Choi, Seong-Soo;Suh, Hong-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.2
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    • pp.109-112
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    • 2002
  • The signal pathways involved in the regulation of AP-1 DNA binding activity in long-term nicotine stimulated bovine adrenal medullary chromaffin (BAMC) cells have not been well characterized. To understand the involvement of second messengers in the regulation of AP-1 DNA binding activity, the present study was designed to define the time-course for inhibition of nicotine-induced responses by cholinergic antagonists, $Ca^{2+}$ and calmodulin (CaM) antagonists, and calcium/calmodulin-dependent protein kinase (CaMK) II inhibitor using electrophoretic mobility shift assay. Nicotine $(10{\mu}M)$ stimulation increased AP-1 DNA binding activity at 24 hr after treatment. Posttreatment with hexamethonium (1 mM) plus atropine $(1{\mu}M)$ (HA), nimodipine $(1{\mu}M),$ or calmidazolium $(1{\mu}M)$ at 0.5, 3, and 6 hr after the nicotine treatment significantly inhibited the AP-1 DNA binding activity increased by long-term nicotine stimulation. However, posttreatment with HA, nimodipine, or calmidazolium at 9 or 12 hr after the nicotine treatment did not affect the nicotine-induced increase of AP-1 DNA binding activity. The pretreatment of BAMC cells with various concentrations of KN-62 inhibited the increase of AP-1 DNA binding activity induced by nicotine in a concentration-dependent manner. KN-62 $(10{\mu}M)$ posttreatment beginning at 0.5, 3, or 6 hr after the nicotine treatment significantly inhibited the increase of AP-1 DNA binding activity. However, KN-62 posttreatment beginning at 9 or 12 hr after the nicotine treatment did not affect the increase of AP-1 DNA binding activity. This study suggested that stimulation (for at least 6 hr) of nicotinic receptors on BAMC cells was necessary for increase of AP-1 DNA binding activity, and activation of $Ca^{2+},$ CaM, and CaMK II up to 6 hr at least seemed to be required for the increase of nicotine-induced AP-1 DNA binding activity.

Suppression of AP-1 Activity by Tanshinone and Cancer Cell Growth Inhibition

  • 박세연;송지성;이덕근;양철학
    • Bulletin of the Korean Chemical Society
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    • v.20 no.8
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    • pp.925-928
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    • 1999
  • The process of transcription is the major point at which gene expression is regulated. The jun and fos families of eukaryotic transcription factor heterodimerize to form complexes capable of binding 5'-TGAGTCA-3'DNA elements (AP-1 binding site). To search for the inhibitors of the jun-fos-DNA complex formation, several natural products extracts were screened and methanol extract of tanshen (the dried roots of Salvia miltiorrhiza Bunge) showed remarkable inhibitory activity. The active compounds of the extracts were purified using re-peated column chromatography and recrystallization. Their structures were identified as tanshinone I and tanshinone IIA. Through the electrophoresis mobility shift assay and cell cytotoxicity test, tanshinone I and tanshinone IIA were identified as inhibitors that suppress not only AP-1 function but also the cell proliferation. Tanshinone I also suppressed the jun-fos-DNA complex formation in TPA-induced NIH 3T3 cells.

Non-histone protein HMGB1 inhibits the repair of damaged DNA by cisplatin in NIH-3T3 murine fibroblasts

  • Yusein-Myashkova, Shazie;Ugrinova, Iva;Pasheva, Evdokia
    • BMB Reports
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    • v.49 no.2
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    • pp.99-104
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    • 2016
  • The nuclear non-histone protein high mobility group box (HMGB) 1 is known to having an inhibitory effect on the repair of DNA damaged by the antitumor drug cisplatin in vitro. To investigate the role of HMGB1 in living cells, we studied the DNA repair of cisplatin damages in mouse fibroblast cell line, NIH-3T3. We evaluated the effect of the post-synthetic acetylation and C-terminal domain of the protein by overexpression of the parental and mutant GFP fused forms of HMGB1. The results revealed that HMGB1 had also an inhibitory effect on the repair of cisplatin damaged DNA in vivo. The silencing of HMGB1 in NIH-3T3 cells increased the cellular DNA repair potential. The increased levels of repair synthesis could be "rescued" and returned to less than normal levels if the knockdown cells were transfected with plasmids encoding HMGB1 and HMGB1 K2A. In this case, the truncated form of HMGB1 also exhibited a slight inhibitory effect.

Identification and Characterization of LHX8 DNA Binding Elements

  • Park, Miree;Jeon, Sanghyun;Jeong, Ji-Hye;Park, Miseon;Lee, Dong-Ryul;Yoon, Tae Ki;Choi, Dong Hee;Choi, Youngsok
    • Development and Reproduction
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    • v.16 no.4
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    • pp.379-384
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    • 2012
  • Lhx8 (LIM homeobox 8) gene encodes a LIM homeodomain transcriptional regulator that is preferentially expressed in germ cells and critical for mammalian folliculogenesis. However, Lhx8 DNA binding sequences are not characterized yet. We aimed to identify and characterize a cis-acting sequence of germ-cell specific transcriptional factor, Lhx8. To identify Lhx8 DNA binding element, Cyclic Amplification of Sequence Target (CAST) Analysis was performed. Electrophoretic Mobility Shift Assay (EMSA) was processed for the binding specificity of Lhx8. Luciferase assay was for the transcriptional activity of Lhx8 through identified DNA binding site. We identified a putative cis-acting sequence, TGATTG as Lhx8 DNA binding element (LBE). In addition, Lhx8 binds to the LBE with high affinity and augments transcriptional activity of luciferase reporter driven by artificial promoter containing the Lhx8 binding element. These findings indicate that Lhx8 directly regulates the transcription of genes containing Lhx8 binding element in oocytes during early folliculogenesis.

Secondary structure analysis of MRA1997 from Mycobacterium tuberculosis and characterization of DNA binding property

  • Kim, Hyo Jung;Lee, Ki-Young;Kim, Yena;Kwon, Ae-Ran;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.2
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    • pp.36-40
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    • 2016
  • MRA1997 is a highly conserved protein from mycobacterial strains. However, no structural and functional information is associated with it. Thus, to obtain details about structure and function of this protein, we have utilized NMR spectroscopy. The recombinant MRA1997 was highly purified and its DNA binding mode was characterized. The tertiary structure of MRA1997 was modeled on the basis of our NMR chemical shift data combined with the webserver CS23D. The binding of MRA1997 with DNA was first monitored by electrophoresis mobility shift assays. The residues involved in DNA binding are identified using NMR chemical shift perturbation experiments. Based on our study, we suggest that MRA1997 interacts with DNA and may play an important role in Mycobacterium tuberculosis physiology.

DNA (Data, Network, AI) Based Intelligent Information Technology (DNA (Data, Network, AI) 기반 지능형 정보 기술)

  • Youn, Joosang;Han, Youn-Hee
    • KIPS Transactions on Computer and Communication Systems
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    • v.9 no.11
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    • pp.247-249
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    • 2020
  • In the era of the 4th industrial revolution, the demand for convergence between ICT technologies is increasing in various fields. Accordingly, a new term that combines data, network, and artificial intelligence technology, DNA (Data, Network, AI) is in use. and has recently become a hot topic. DNA has various potential technology to be able to develop intelligent application in the real world. Therefore, this paper introduces the reviewed papers on the service image placement mechanism based on the logical fog network, the mobility support scheme based on machine learning for Industrial wireless sensor network, the prediction of the following BCI performance by means of spectral EEG characteristics, the warning classification method based on artificial neural network using topics of source code and natural language processing model for data visualization interaction with chatbot, related on DNA technology.

Temperature-dependent DNA binding of DicA protein in vivo and in vitro (In vivo와 in vitro에서 DicA 단백질의 온도 의존적 DNA 결합)

  • Lee, Yonho;Yun, Sang Hoon;Lim, Heon M.
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.181-190
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    • 2019
  • In Escherichia coli, DicA protein is involved in cell division control. DicA protein is known to bind DNA better at $25^{\circ}C$ than at $37^{\circ}C$. However, the molecular cause of the temperature dependent binding is not clear. In this study, we investigated how DicA binds DNA and why its DNA binding activity depends on temperature. An unique in vivo DNA binding assay developed in this laboratory showed that unlike the homologous proteins such as RovA or SlyA, DicA uses its N-terminal domain for DNA binding. The in vivo DNA binding assay of DicA also demonstrated that the temperature-dependent DNA binding activity does not come from Cnu or H-NS that is known to bind DNA better at $25^{\circ}C$ than at $37^{\circ}C$. Electrophoretic Mobility Shift Assay (EMSA), when performed with purified DicA protein, did not show temperature-dependent DicA binding activity. However when EMSA was performed with crude protein from WT E. coli cells, temperature-dependent DicA binding activity was observed, suggesting that there is a factor(s) that confers temperature DNA binding activity of DicA in vivo.

Epidermal Growth Factor Receptor Gene Polymorphisms and Gastric Cancer in Iran

  • Abediankenari, Saeid;Jeivad, Fereshteh
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3187-3190
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    • 2013
  • Background: Epidermal growth factor receptor (EGFR) is a transmembrane receptor which contributes to many processes involved in cell survival, proliferation and inhibits apoptosis, that may lead to cancer development. Gastric cancer is one of the most common diseases of digestive system that has low 5-year-survival. The aim of this research was to determine the significance of EGFR tyrosine kinase domain gene polymorphisms in gastric cancer in Iran. Materials and Methods: In the present study, 83 patients with gastric cancer and 40 normal subjects were investigated for EGFR gene polymorphisms in exons 18-21 by PCR-SSCP. Then, DNA sequencing was conducted for different mobility shift bands. Finally the data were statistically analyzed using the chi-2 test and the SPSSver.16 program. Results: Exon 18 of EGFR gene showed three different bands in SSCP pattern and DNA sequencing displayed one mutation. SSCP pattern of Exons 19 and 21 did not show different migration bands. Exon 20 of EGFR gene revealed multiple migrate bands in SSCP pattern. DNA sequencing displayed 2 mutations in this exon: one mutation was caused amino acid change and another mutation was silent. Conclusion: It may be that EGFR tyrosine kinase gene polymorphisms differ between populations and screening could be useful in gastric cancer patients who might benefit from tyrosine kinase inhibitor therapy.

Intrinsic bent DNA colocalizes with the sequence involved in the Nd-sD mutation in the Bombyx mori fibroin light chain gene

  • Barbosa, Joice Felipes;Bravo, Juliana Pereira;Takeda, Karen Izumi;Zanatta, Daniela Bertolini;Silva, Jose Luis Da Conceicao;Balani, Valerio Americo;Fiorini, Adriana;Fernandez, Maria Aparecida
    • BMB Reports
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    • v.41 no.5
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    • pp.394-399
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    • 2008
  • Multiple sequence alignments of the Bombyx mori fibroin light chain gene (fib-L) from hybrids and from Chinese and Japanese strains demonstrated that 51.6% of the fib-L third intron is conserved. One of these conserved segments, 41 bp long, contains the sequence CGTTATTATACATATT, which is duplicated in the B. mori Nd-$s^D$ mutant. In the present work, electrophoretic mobility assays and computational analyses revealed a major peak of intrinsic bent DNA within the segment that undergoes breakage in the previously-described Nd-$s^D$ mutation. This result suggested that this intrinsically-curved region might mediate DNA cleavage and enhance recombination events in the third intron of the Bombyx mori fib-L gene.