• Title/Summary/Keyword: DNA interaction

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Dephosphorylation of DBC1 by Protein Phosphatase 4 Is Important for p53-Mediated Cellular Functions

  • Lee, Jihye;Adelmant, Guillaume;Marto, Jarrod A.;Lee, Dong-Hyun
    • Molecules and Cells
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    • v.38 no.8
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    • pp.697-704
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    • 2015
  • Deleted in breast cancer-1 (DBC1) contributes to the regulation of cell survival and apoptosis. Recent studies demonstrated that DBC is phosphorylated at Thr454 by ATM/ATR kinases in response to DNA damage, which is a critical event for p53 activation and apoptosis. However, how DBC1 phosphorylation is regulated has not been studied. Here we show that protein phosphatase 4 (PP4) dephosphorylates DBC1, regulating its role in DNA damage response. PP4R2, a regulatory subunit of PP4, mediates the interaction between DBC1 and PP4C, a catalytic subunit. PP4C efficiently dephosphorylates pThr454 on DBC1 in vitro, and the depletion of PP4C/PP4R2 in cells alters the kinetics of DBC1 phosphorylation and p53 activation, and increases apoptosis in response to DNA damage, which are compatible with the expression of the phosphomimetic DBC-1 mutant (T454E). These suggest that the PP4-mediated dephosphorylation of DBC1 is necessary for efficient damage responses in cells.

Electrochemical Detection of $17{\beta}-estradiol$ by using DNA Aptamer Immobilized Nanowell Gold Electrodes

  • Kim, Yeon-Seok;Jung, Ho-Sup;Lee, Hea-Yeon;Kawai, Tomoji;Gu, Man-Bock
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.88-92
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    • 2005
  • Aptamer is the single-stranded oligonucleotide which binds to various target molecules such as proteins, peptides, lipids and small organic molecules with high affinity and specificity. DNA aptamers specific for the $17{\beta}-estradiol$ were selected by SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random DNA library. These DNA aptamers have a high affinity to $17{\beta}-estradiol$ as an endocrine disrupting chemical. Nanowell and $200{\mu}m$ gold electrode were used as substrate for DNA aptamer immobilization and electrochemical analysis. Especially, nanowell gold electrode was fabricated by e-beam lithography. The size of single nanowell is 130nm and 40,000 nanowells were deposited on one gold electrode. The immobilization method was based on the interaction between the biotinylated aptamer and streptavidin deposited on gold electrode previously. Immobilization procedure was optimized by surface plasma resonance (SPR) and electrochemical analysis. After the immobilization of DNA aptamer on streptavidin modified gold electrode, $17{\beta}-estradiol$ solution was treated on aptamer immobilized gold electrode. The current of gold electrode was decreased by the binding of $17{\beta}-estradiol$ to DNA aptamer immobilized on gold electrode. However, in negative control experiments of 1-aminoanthraquinone and 2-methoxynaphthalene, the current was rarely decreased. And more sensitive data was obtained from nanowell gold electrode comparing with $200{\mu}m$ gold electrode.

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Applications of DNA Microarray in Disease Diagnostics

  • Yoo, Seung-Min;Choi, Jong-Hyun;Lee, Sang-Yup;Yoo, Nae-Choon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.635-646
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    • 2009
  • Rapid and accurate diagnosis of diseases is very important for appropriate treatment of patients. Recent advances in molecular-level interaction and detection technologies are upgrading the clinical diagnostics by providing new ways of diagnosis, with higher speed and accuracy. In particular, DNA microarrays can be efficiently used in clinical diagnostics which span from discovery of diseaserelevant genes to diagnosis using its biomarkers. Diagnostic DNA microarrays have been used for genotyping and determination of disease-relevant genes or agents causing diseases, mutation analysis, screening of single nucleotide polymorphisms (SNPs), detection of chromosome abnormalities, and global determination of posttranslational modification. The performance of DNA-microarray-based diagnosis is continuously improving by the integration of other tools. Thus, DNA microarrays will play a central role in clinical diagnostics and will become a gold standard method for disease diagnosis. In this paper, various applications of DNA microarrays in disease diagnosis are reviewed. Special effort was made to cover the information disclosed in the patents so that recent trends and missing applications can be revealed.

Role of cAMP, EGF, IGF-I and Protein Phosphorylation in Mammary Development I. Effect of EGF, IGF-I and Photoreactive Cyclic AMP on DNA Synthesis of Mammary Epithelial Cell (유선발달에 있어서 cAMP, EGF, IGF-I 및 단백질 인산화 작용의 역할 I. EGF, IGF-I 및 Photoreactive Cyclic AMP가 유선상피세포의 DNA합성에 미치는 효과)

  • 여인서;박춘근;홍병주
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.49-56
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    • 1993
  • Mouse mammary epithelial cells(NMuMG) were plated onto 24 well phates(100,000 cells/well), in DMEM supplemented with 10% fetal calf serum. After serum starvation for 24 hours, EGF)0~100ng/ml) was added simultaneously with IGF-I(10ng/ml), 1$\mu$M photoreactive cAMP(4,5-dimethoxy-2-nitrobenzyl adenosine-3',5' cyclic monophosphate, DMNB) or IGF-I plus DMNB. After 2 hours, the cells were expposed to UV light(300nm, 3 second pulse0 in order to activate DMNB which induces a rapid transient increase in intracellular cAMP upon UV irradiation. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA(1 hour pulse with 1$\mu$Ci/ml, 18~19 hours after UV exposure). Without IGF-I or DMNB, EGF(10 or 100ng/ml) increased DNA synthesis from 8,362 dpm/well in control to 16,345 or 18,684 dpm/well with EGF(pooled SE=1,239 dpm/well, P<0.05). IGF-I or IGF-I plus DMNB alone increased DNA synthesis from 8,362 dpm/well in control to 17,307 or 20,427 dpm/well, respectively(P<0.05). Addition of IGF-I, DMNB or IGF-I plus DMNB into 0~100ng/ml EGF did not significantly change the shape of dose response curve of EGF alone. In other experiment, EGF or IGF-I plus DMNB into 10ng/ml EGF group exhibited interaction effect in DNAsynthesis [EGF(10ng/ml)=18,497; IGF-I+EGF=22,837; DMNB+EGF=20,658 ; IGF-I+DMNB+EGF=29,658, pooled SE=1,055, P<0.05]. These results indicate that simultaneous activation of EGF, IGF-I and intracellular cAMP interact in DNA synthesis of mouse mammary epithelial cells.

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Characterization of a New Gene Resistant to Alkylating Agents and 3-Aminobenzamide When Knocked Out in Fission Yeast (분열형 효모에서 유전자 결실에 의해 알킬화제와 3-AMINOBENZAMIDE에 저항성을 나타내는 새로운 유전자의 특성 분석)

  • 박종군;차재영;황성진;박세근;김미영;백성민;최인순;이정섭
    • Journal of Life Science
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    • v.12 no.2
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    • pp.219-225
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    • 2002
  • The organization of eukayotic chromatin into specific conformation that are associated with transcription, replication, reapir and other nuclear processes are achieved via a series of DNA-protein interaction. These interactions are mediated by a range of DNA-binding domains such as SAP domain et at. By searching S. pombe genomic DNA database, we have found a gene named SAPuvs (SAP UV Sensitive) whose amino acid sequence is in part similar to SAP domain of Arabidopsis poly (ADP-ribose) polymerase and Ku7O. Knock-out cell of S. pombe SAPuvs gene was constructed using Ura4 as a selection marker. Survival analysis of knock-out cell indicated that treatment with UV significantly reduces the survival compared to wild type cell. Potentiation of MMS-induced cytotoxicity by 3AB post-treatment was observed in wild type cells, but not in knock-out cells. These data suggested that the protein encoded by SAPuvs gene is associated with chromatin reorganization during DNA repair.

Polymorphisms of XRCC1 and XRCC2 DNA Repair Genes and Interaction with Environmental Factors Influence the Risk of Nasopharyngeal Carcinoma in Northeast India

  • Singh, Seram Anil;Ghosh, Sankar Kumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.2811-2819
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    • 2016
  • Multiple genetic and environmental factors have been reported to play key role in the development of nasopharyngeal carcinoma (NPC). Here, we investigated interactions of XRCC1 Arg399Gln and XRCC2 Arg188His polymorphisms and environmental factors in modulating susceptibility to NPC in Northeast India. One-hundred NPC patients, 90 first-degree relatives of patients and 120 controls were enrolled in the study. XRCC1 Arg399Gln and XRCC2 Arg188His polymorphisms were determined using PCR-RFLP, and the results were confirmed by DNA sequencing. Logistic regression (LR) and multifactor dimensionality reduction (MDR) approaches were applied for statistical analysis. The XRCC1 Gln/Gln genotype showed increased risk (OR=2.76; P<0.024) of NPC. However, individuals with both XRCC1 and XRCC2 polymorphic variants had 3.2 fold elevated risk (P<0.041). An enhanced risk of NPC was also observed in smoked meat (OR=4.07; P=0.004) and fermented fish consumers (OR=4.34, P=0.001), and tobacco-betel quid chewers (OR=7.00; P=0.0001) carrying XRCC1 polymorphic variants. However, smokers carrying defective XRCC1 gene showed the highest risk (OR = 7.47; P<0.0001). On MDR analysis, the best model for NPC risk was the five-factor model combination of XRCC1 variant genotype, fermented fish, smoked meat, smoking and chewing (CVC=10/10; TBA=0.636; P<0.0001); whereas in interaction entropy graphs, smoked meat and tobacco chewing showed synergistic interactions with XRCC1. These findings suggest that interaction of genetic and environmental factors might increase susceptibility to NPC in Northeast Indian populations.

Identification of the Interaction between Insulin-like Growth Factor Binding Protein-4 (IGFBP-4) and Heterogeneous Nuclear Ribonucleoprotein L (hnRNP L) (IGF결합 단백질-4(IGFBP-4)와 이질 핵 리보핵산단백질 L (hnRNP L)의 상호결합의 식별)

  • Choi, Mieyoung
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1311-1316
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    • 2013
  • Heterogeneous nuclear ribonucleoprotein L (hnRNP L) is a major pre-mRNA binding protein and it is an abundant nuclear protein that shuttles between the nucleus and the cytoplasm. hnRNP L is known to be related to many cellular processes, including chromatin modification, pre-mRNA splicing, mRNA export of intronless genes, internal ribosomal entry site (IRES)-mediated translation, mRNA stability, and spermatogenesis. In order to identify the cellular proteins interacting with hnRNP L, this study performed a yeast two-hybrid screening, using a human liver cDNA library. The study identified insulin-like growth factor binding protein-4 (IGFBP-4) as a novel interaction partner of hnRNP L in the human liver. It then discovered, for the first time, that hnRNP L interacts specifically with IGFBP-4 in a yeast two-hybrid system. The authenticity of this two-hybrid interaction of hnRNP L and IGFBP-4 was confirmed by an in vitro pull-down assay.

Vitamin B12 Model Complexes: Synthesis and Characterization of Thiocyanato Cobaloximes and Thiocyanato Bridged Dicobaloximes of O-donor Ligands: DNA Binding and Antimicrobial Activity (비타민 B12 모델 착물: O-주개 리간드인 Thiocyanato Cobaloximes 및 Thiocyanato로 연결된 Dicobaloximes의 합성 및 특성규명: DNA 결합 및 향균 활성)

  • Mustafa, Bakheit;Satyanarayana, S.
    • Journal of the Korean Chemical Society
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    • v.54 no.6
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    • pp.687-695
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    • 2010
  • Complexes of thiocyanato(L)cobaloximes where L is urea, acetamide, semicrabazide and formamide were synthesized and characterized. The reaction of thiocyanato (L) cobaloximes (SCNCo$(DH)_2$(L)) with benzyl (aquo) cobaloxime $PhCH_2Co(DH)_2(OH_2)$ was found to produce a series of thiocyanato bridged dicobaloximes of a general formula of $PhCH_2Co(DH)_2SCNCo(DH_2)(L)$. Evidence for formulation as dicobaloximes containing thiocyanato ligand bridges was obtained from infrared data which show $20-45cm^{-1}$ increase in vCN upon formation of the dicobaloxime from the corresponding terminal thiocyanocobaloxime (SCNCo$(DH)_2$(L)). Further characterization of these two series was done on the basis of ($^1H$,$^{13}C$)NMR, LCMS and elemental analysis. Anti-microbial activity of thiocyanato(L)cobaloximes and thiocyanato bridged dicobaloximes were screened against E. Coli. The DNA-binding behaviors of both monomers and dimers were investigated by spectroscopic methods and viscosity measurements. The results indicated that the dimer complexes bind with calf-thymus DNA in an intercalative mode via the terminal benzyl ring into the base pairs of DNA. It was observed that the monomer complexes did not interact with DNA. Fluorescence spectra for the interaction between thiocyanato bridged dicobaloximes and DNA were also studied.

Cochleicola gelatinilyticus gen. nov., sp. nov., Isolated from a Marine Gastropod, Reichia luteostoma

  • Shin, Su-Kyoung;Kim, Eunji;Choi, Sungmi;Yi, Hana
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1439-1445
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    • 2016
  • A yellow, rod-shaped, non-motile, gram-negative, and strictly aerobic bacterial strain, designated LPB0005T, was isolated from a marine gastropod, Reichia luteostoma. Here the genome sequence was determined, which comprised 3,395,737 bp with 2,962 protein-coding genes. The DNA G+C content was 36.3 mol%. The 16S rRNA gene sequence analysis indicated that the isolate represents a novel genus and species in the family Flavobacteriaceae, with relatively low sequence similarities to other closely related genera. The isolate showed chemotaxonomic properties within the range reported for the family Flavobacteriaceae, but possesses many physiological and biochemical characteristics that distinguished it from species in the closely related genera Ulvibacter, Jejudonia, and Aureitalea. Based on phylogenetic, phenotypic, and genomic analyses, strain LPB0005T represents a novel genus and species, for which the name Cochleicola gelatinilyticus gen. nov., sp. nov. is proposed. The type strain is LPB0005T (= KACC 18693T = JCM 31218T).

Optical sensitivity of DNA-dispersed single-walled carbon nanotubes within cement composites under mechanical load

  • Kim, Jin Hee;Rhee, Inkyu;Jung, Yong Chae;Ha, Sumin;Kim, Yoong Ahm
    • Carbon letters
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    • v.24
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    • pp.90-96
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    • 2017
  • We demonstrated the sensitivity of optically active single-walled carbon nanotubes (SWCNTs) with a diameter below 1 nm that were homogeneously dispersed in cement composites under a mechanical load. Deoxyribonucleic acid (DNA) was selected as the dispersing agent to achieve a homogeneous dispersion of SWCNTs in an aqueous solution, and the dispersion state of the SWCNTs were characterized using various optical tools. It was found that the addition of a large amount of DNA prohibited the structural evolution of calcium hydroxide and calcium silicate hydrate. Based on the in-situ Raman and X-ray diffraction studies, it was evident that hydrophilic functional groups within the DNA strongly retarded the hydration reaction. The optimum amount of DNA with respect to the cement was found to be 0.05 wt%. The strong Raman signals coming from the SWCNTs entrapped in the cement composites enabled us to understand their dispersion state within the cement as well as their interfacial interaction. The G and G' bands of the SWCNTs sensitively varied under mechanical compression. Our results indicate that an extremely small amount of SWCNTs can be used as an optical strain sensor if they are homogeneously dispersed within cement composites.