• Title/Summary/Keyword: DNA 결합

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Cloning of hnRNP E1 cDNA via yeast two-hybrid system and a study on protein-protein interaction between hnRNP E1 and hnRNP K (이스트 two-hybrid 시스템을 이용한 hnRNP E1 cDNA의 클로닝과 hnRNP E1-hnRNP K 상호결합에 대한 연구)

  • Choi, Mie-Young
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.9 no.6
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    • pp.1795-1799
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    • 2008
  • The heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a component of hnRNP complexes. This protein binds strongly to cytidine-rich RNA/DNA sequences. It is a nucleocytoplasmic shuttling protein. To investigate the functions of hnRNP K, I searched for hnRNP K-interacting proteins in HeLa cDNA library using a yeast two-hybrid screening system. One of the cDNA clones is identical to human hnRNP E1 (poly(rC) binding protein 1) cDNA (GenBank accession number XM_031585). In this study, hnRNP K is shown to specifically interact with hnRNP E1 in yeast two-hybrid system and in vitro biochemical assay.

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.

An Error Detection and Repair on DNA Duplicate Structure (DNA 이중나선구조에서의 오류 검출 및 복구)

  • Kim, Soke-Hwan;Hur, Chang-Wu
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.15 no.11
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    • pp.2500-2504
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    • 2011
  • Living organisms are composed of cells that can replicate themselves through growth, division packed with tons capacity. On DNA mutations, ie mutations in the offspring's survival and reproduction can be held against you, and packed with tons ambivalence that could benefit. In this study, the DNA double helix is used as a template for replication, we first separated into single strands of the double helix must be opened Combining the double helix portion of the location of errors in the bond provides a way to find and repair.

DNA 염기의 토토머화에 의한 염기쌍 변이 연구: 염기쌍의 전이 돌연변이를 중심으로

  • Lee, Yeon-Hui;Lee, Min-Jun;Sin, Seok-Min
    • Proceeding of EDISON Challenge
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    • 2015.03a
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    • pp.99-104
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    • 2015
  • DNA 복제는 굉장한 정확도를 가지고 이루어진다. 하지만 내, 외부적인 여러 요인으로 돌연변이가 일어나기도 한다. 그 중 토토머화에 의해 치환 돌연변이가 일어난다는 가설은 오래전부터 그 가능성이 논의되어 왔고, 직접적인 증거를 찾으려는 노력도 있어 왔다. 토토머화에 의하여 DNA염기가 다른 형으로 변하면 왓슨-크릭 염기쌍 (아데닌-타이민, 시토신-구아닌이 수소결합한 염기쌍)이 아닌 다른 염기쌍이 생성될 수 있다. 이 염기쌍이 기준이 되어 DNA가 복제되기 때문에 결과적으로 전이 돌연변이(transition), 혹은 전환 돌연변이(transversion)된 염기쌍이 생성될 수 있다. 우리는 이런 사례 중에 염기쌍의 전이 돌연변이를 중심으로 연구하고자 한다. A-T염기쌍중 하나가 지배적인 아민형(amine form)이 아니라 이민형(imine form)으로 존재할 때 아민형과는 다른 수소결합이 가능해지며 보통 잘 생성되지 않는 A-C, G-T결합이 생성될 수 있다. 이후 그 염기쌍이 기준이 되어 DNA가 복제될 때에는 왓슨-크릭 염기쌍이 주로 생성되어 A-T염기쌍이 G-C 염기쌍으로 치환되는 DNA가닥이 생기게 된다. 우리는 이러한 과정을 에너지와 반응속도 측면에 집중하여 분석해보았다. 계산 결과, $A-C^*$, $A^*-C$, $G-C^*$, $G^*-C$의 염기쌍 생성이 왓슨-크릭 염기쌍과 비슷하거나 더 큰 정도로 에너지 면에서 유리하였으며 대부분의 돌연변이가 에너지 측면에서 보았을 때 $A-C^*$ 염기쌍을 통하여 생김을 알 수 있었다. 계산된 값은 약 $10^{-6}$ 정도의 빈도를 보였다.

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Effect of Ginseng Extracts on the Binding to DNA of Benzo(a)pyrene Metabolites in uitro in Rats (DNA와 Benzo(a)pyrene 대사물질 결합형성에 미치는 인삼 추출물의 영향)

  • 박진규;고지훈
    • Journal of Ginseng Research
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    • v.13 no.1
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    • pp.37-41
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    • 1989
  • Reactive metabolites generated by benzo(a)pyrene(BP) monooxygenase(AHH) interact with nucleophiles in DNA and cause mutation and carcinogenesis. We studied the effect of Panax ginseng C.A. Meyer, which induce epoxide hydratase(EH) activity without concomitant induction of AHH activity, on the binding of BP metabolites to DNA in uitro in Sprague Dawley rats. DNA-BP metabolite adducts can be resolved into at least five distinct peaks by elution of a Sephadex LH-20 column with a water methanol gradieNt. These peaks are arbitrarily designated A(most polar) through I(least polar). Of the 5 peaks tentatively assigned to 7,8 biol-9,10-oxide(A),7,8·oxide(B),4,5-oxide(C), and further metabolites of 9-OH-BP(D & E), peaks A, C, D, and I were reduced to 70, 85, 80, and 30% of controls, respectively, and there was no significant change in peak B. In connection with this DNA binding study, BP metabolizing enzymes including AHH, EH, demethylase(DM) activity and cyt. P-450 contents were also investigated in order to compare the BP treated control with ginseng and BP treated test groups. The results showed that the EH activity was increased by 139% over the BP control, the Cyt. P-450 content was increased by 180% over the control value, and DM and AHH activities were also increased to some degree for the BP test group, but there was no significant effect of the ginseng treatment.

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H-NS binding on dicA promoter DNA inhibits dicA gene expression (dicA promoter DNA에 붙는 H-NS 단백질에 의한 dicA 유전자의 발현 조절)

  • Yun, Sang Hoon;Lee, Yonho;Lim, Heon M.
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.191-198
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    • 2019
  • H-NS binds to promoter DNA and works as a general transcription silencer. DicA protein, by binding to the promoter DNA of dicA, activates dicA expression and at the same time inhibits expression of dicF and dicB, thus, exerting cell division control in Escherichia coli. H-NS complexed with a nucleoid protein Cnu was known to be involved in dicA expression. However, the exact nature of H-NS binding to dicA promoter DNA and the consequences of H-NS binding in expression of dicA is not clear. In this study, we explored the DNA binding activity of H-NS on the promoter DNA of dicA and found that H-NS binding occurs exclusively to the dicA promoter DNA. We never observed, however, H-NS binding at the vicinity of the dicA promoter. Temperature dependent oligomerization of H-NS was observed during DNA binding and the Cnu protein enhances the oligomerization process of H-NS binding. In vivo measurement of dicA expression in an hns deleted strain showed that dicA expression increased. These results demonstrated that H-NS binds specifically to dicA promoter DNA and functions as a transcription silencer.

CheY-OmpR Hybrid Protein Acting on the Osmoregulatory System (CheY-OmpR 혼성 단백질의 삼투조절효과)

  • 고민수;박찬규
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.118-124
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    • 1997
  • In the previous study(6), we constructed the CheY-OmpR hybrid, Chp, which affects the expressions of ompF and om pC genes. Here we further characterize these effects and present the regulatory mechanism based on in vivo and in vitro data. Although Chp retained the sequence-specific DNA-binding ability, it was not possible to enhance transcriptional activity, suggesting that it may act as a competitive inhibitor to OmpR. The DNA-binding affinity of Chp was not modulated by phosphorylation of its Che Y portion. Chp was able to increase ompR transcription. FurthemlOre, it was found that the wild-type OmpR also exerts the same effect, which is also eOlltrolled by changes in medium osmolarity and in EnvZ activity.

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Protonation State of Norfloxacin and Their Interaction with DNA (pH에 따른 Norfloxacin의 형태 및 DNA와의 상호작용에 관한 연구)

  • Yeo, Jeong-Ah;Son, Gwan-Su;Kim, Jong-Moon;Moon, Hyung-Rang;Jun, Eun-Duk;Cho, Tae-Sub
    • Journal of the Korean Chemical Society
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    • v.44 no.1
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    • pp.4-9
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    • 2000
  • We investigated the self-association and protonation state of norfloxacin, a member of quinokme antibiotics, using electric absorption and potentiom,etric titration. Both nitrogen at the piperazine ring and carboxylic acid were protonated at a low pH (cationic norfloxacin), and deprotonated at a high pH (anionic norfloxacin). In the intermediate pH range, a neutral species was dominant with the possibility of forming a zwitter ion. We also observed that nortloxacin molecules can be sracked to form a dimer at an intermediate pH, The equilibrium constant of the norfloxacin-DNA complex formation, which was measured by Stem-Volmermethod, increases as the pH of the system is lowered. This observation indicates that it is the cationic nortloxacin that forms a complex with DNA among various norfloxacin species in aqueous solution.

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Application of Structure-Switching Signaling Aptamers in DNA computing (DNA 컴퓨팅에서의 앱타머 구조 변환 활용 방안)

  • 김수동;장병탁
    • Proceedings of the Korean Information Science Society Conference
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    • 2003.10b
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    • pp.838-840
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    • 2003
  • 특정 단백질과 특이적으로 결합하는 핵산인 앱타머 (aptamer) 의 존재는, DNA 기반 컴퓨팅과 단백질 기반 컴퓨팅 사이에서 가교 역할을 할 수 있다는 가능성을 고려할 때 주목할 만하다. 본고에서는 전통적인 DNA 기반 컴퓨팅 방법론의 확장으로서, 앱타머 구조 변환의 활용 방안을 제안하였다.

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The Role of DNA Binding Domain in hHSF1 through Redox State (산화환원에 따른 hHSF1의 DNA binding domain의 역할)

  • Kim, Sol;Hwang, Yun-Jeong;Kim, Hee-Eun;Lu, Ming;Kim, An-D-Re;Moon, Ji-Young;Kang, Ho-Sung;Park, Jang-Su
    • Journal of Life Science
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    • v.16 no.6
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    • pp.1052-1059
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    • 2006
  • The heat shock response is induced by environmental stress, pathophysiological state and non-stress conditions and wide spread from bacteria to human. Although translations of most proteins are stopped under a heat shock response, heat shock proteins (HSPs) are produced to protect cell from stress. When heat shock response is induced, conformation of HSF1 was changed from monomer to trimer and HSF1 specifically binds to DNA, which was called a heat shock element(HSE) within the promoter of the heat shock genes. Human HSF1(hHSFl) contains five cysteine(Cys) residues. A thiol group(R-SH) of Cys is a strong nucleophile, the most readily oxidized and nitrosylated in amino acid chain. This consideration suggests that Cys residues may regulate the change of conformation and the activity of hHSF1 through a redox-dependent thiol/disulfide exchange reaction. We want to construct role of five Cys residues of hHSF by redox reagents. According to two studies, Cys residues are related to trimer formation of hHSF1. In this study, we want to demonstrate the correlation between structural change and DNA-binding activity of HSF1 through forming disulfide bond and trimerization. In this results, we could deduce that DNA binding activity of DNA binding domain wasn't affected by redox for always expose outside to easily bind to DNA. DNA binding activity of wild-type HSF's DNA binding domain was affected by conformational change, as conformational structure change (trimerization) caused DNA binding domain.