• Title/Summary/Keyword: zinc fingers

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Structural Analyses of Zinc Finger Domains for Specific Interactions with DNA

  • Eom, Ki Seong;Cheong, Jin Sung;Lee, Seung Jae
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2019-2029
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    • 2016
  • Zinc finger proteins are among the most extensively applied metalloproteins in the field of biotechnology owing to their unique structural and functional aspects as transcriptional and translational regulators. The classical zinc fingers are the largest family of zinc proteins and they provide critical roles in physiological systems from prokaryotes to eukaryotes. Two cysteine and two histidine residues ($Cys_2His_2$) coordinate to the zinc ion for the structural functions to generate a ${\beta}{\beta}{\alpha}$ fold, and this secondary structure supports specific interactions with their binding partners, including DNA, RNA, lipids, proteins, and small molecules. In this account, the structural similarity and differences of well-known $Cys_2His_2$-type zinc fingers such as zinc interaction factor 268 (ZIF268), transcription factor IIIA (TFIIIA), GAGA, and Ros will be explained. These proteins perform their specific roles in species from archaea to eukaryotes and they show significant structural similarity; however, their aligned amino acids present low sequence homology. These zinc finger proteins have different numbers of domains for their structural roles to maintain biological progress through transcriptional regulations from exogenous stresses. The superimposed structures of these finger domains provide interesting details when these fingers are applied to specific gene binding and editing. The structural information in this study will aid in the selection of unique types of zinc finger applications in vivo and in vitro approaches, because biophysical backgrounds including complex structures and binding affinities aid in the protein design area.

ZNF435, a Novel Human SCAN-containing Zinc Finger Protein, Inhibits AP-1-mediated Transcriptional Activation

  • Gu, Xing;Zheng, Mei;Fei, Xiangwei;Yang, Zhenxing;Li, Fan;Ji, Chaoneng;Xie, Yi;Mao, Yumin
    • Molecules and Cells
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    • v.23 no.3
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    • pp.316-322
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    • 2007
  • Zinc finger transcription factor genes are a significant fraction of the genes in the vertebrate genome. Here we report the isolation and characterization of a human zinc finger-containing gene, ZNF435, from a fetal brain cDNA library. ZNF435 cDNA is 1290 base pairs in length and contains an open reading frame encoding 349 amino acids with four C2H2-type zinc fingers at its carboxyl terminus and a SCAN motif at its amino terminus. RT-PCR results showed that ZNF435 was expressed in all tested tissues. A ZNF435-GFP fusion protein was located in the nucleus and the four zinc fingers acted as nuclear localization signals (NLSs). ZNF435 was found to be capable of homo-association, and this effect was independent of its zinc fingers. Furthermore, ZNF435 proved to be a transcription repressor as its overexpression in AD293 cells inhibited the transcriptional activities of AP-1.

Engineering and Application of Zinc Finger Proteins and TALEs for Biomedical Research

  • Kim, Moon-Soo;Kini, Anu Ganesh
    • Molecules and Cells
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    • v.40 no.8
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    • pp.533-541
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    • 2017
  • Engineered DNA-binding domains provide a powerful technology for numerous biomedical studies due to their ability to recognize specific DNA sequences. Zinc fingers (ZF) are one of the most common DNA-binding domains and have been extensively studied for a variety of applications, such as gene regulation, genome engineering and diagnostics. Another novel DNA-binding domain known as a transcriptional activator-like effector (TALE) has been more recently discovered, which has a previously undescribed DNA-binding mode. Due to their modular architecture and flexibility, TALEs have been rapidly developed into artificial gene targeting reagents. Here, we describe the methods used to design these DNA-binding proteins and their key applications in biomedical research.

A Splice Variant of the C2H2-Type Zinc Finger Protein, ZNF268s, Regulates NF-κB Activation by TNF-α

  • Chun, Jung Nyeo;Song, In Sung;Kang, Dong-Hoon;Song, Hye Jin;Kim, Hye In;Suh, Ja Won;Lee, Kong Ju;Kim, Jaesang;Won, Sang
    • Molecules and Cells
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    • v.26 no.2
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    • pp.175-180
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    • 2008
  • $I{\kappa}B$ kinase (IKK), the pivotal kinase in signal-dependent activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$), is composed of multiple protein components, including IKK ${\alpha}/{\beta}/{\gamma}$ core subunits. To investigate the regulation of the IKK complex, we immunoaffinity purified the IKK complex, and by MALDI-TOF mass spectrometry identified a splice variant of zinc finger protein 268 (ZNF268) as a novel IKKinteracting protein. Both the full-length and the spliced form of the ZNF268 protein were detected in a variety of mammalian tissues and cell lines. The genes were cloned and expressed by in vitro transcription/translation. Several deletion derivatives, such as KRAB domain (KRAB) on its own, the KRAB/spacer/4-zinc fingers (zF4), and the spacer/4-zinc fingers (zS4), were ectopically expressed in mammalian cells and exhibited had different subcellular locations. The KRAB-containing mutants were restricted to the nucleus, while zS4 was localized in the cytosol. TNF-${\alpha}$-induced NF-${\kappa}B$ activation was examined using these mutants and only zS4 was found to stimulate activation. Collectively, the results indicate that a spliced form of ZNF268 lacking the KRAB domain is located in the cytosol, where it seems to play a role in TNF-${\alpha}$-induced NF-${\kappa}B$ activation by interacting with the IKK complex.

Development of Anti-Fingers Printed EGI Steel Sheet With High Corrosion Resistance (고내식성 내지문 EGI 강판 개발)

  • 김형준
    • Journal of the Korean institute of surface engineering
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    • v.26 no.6
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    • pp.307-315
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    • 1993
  • A study on a new anti-finger printed EGI steel sheet with high corrosion resistance has been carried out to meet the recent requirement of high quality and performance of the pure zinc electrogalvanized steel sheet. The substrate was the pure zinc electrogalvanized sheet with the metallic coating of 20g/$\m^2$. The two differ-ent processes for inorganic(chromating) and organic(resin) coating were applied. One was a two coat/two bake type to separately treat chromating and resin coating which is now widely used. The other was a one coat/one bake type to simultaneously treat them which is newly developed in this study. The solution for the one coat/one bake type was an aqua-base coating agents which was composed of inorganic and organic components. The new anti-finger printed EGI steel sheet with the Cr and resin coating weight of 13mg/$\m^2$ and 800mg/$\m^2$, respectively shows the superior corrosion resistance besides the good paintability, formability fingerprint resistance and earth characteristics properties.

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Isolation and Characterization of Pathogen-Inducible Putative Zinc Finger DNA Binding Protein from Hot Pepper Capsicum annuum L.

  • Oh, Sang-Keun;Park, Jeong-Mee;Jung, Young-Hee;Lee, Sanghyeob;Kim, Soo-Yong;Eunsook Chung;Yi, So-Young;Kim, Young-Cheol;Seung, Eun-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.79.2-80
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    • 2003
  • To better understand plant defense responses against pathogen attack, we identified the transcription factor-encoding genes in the hot pepper Capsicum annuum that show altered expression patterns during the hypersensitive response raised by challenge with bacterial pathogens. One of these genes, Ca1244, was characterized further. This gene encodes a plant-specific Type IIIA - zinc finger protein that contains two Cys$_2$His$_2$zinc fingers. Ca1244 expression is rapidly and specifically induced when pepper plants are challenged with bacterial pathogens to which they are resistant. In contrast, challenge with a pathogen to which the plants are susceptible only generates weak Ca1244 expression. Ca1244 expression is also strongly induced in pepper leaves by the exogenous application of ethephon, an ethylene releasing compound. Whereas, salicylic acid and methyl jasmonate had moderate effects. Pepper protoplasts expressing a Ca1244-smGFP fusion protein showed Ca1244 localizes in the nucleus. Transgenic tobacco plants overexpressing Ca1244 driven by the CaMV 355 promoter show increased resistance to challenge with a tobacco-specific bacterial pathogen. These plants also showed constitutive upregulation of the expression of multiple defense-related genes. These observations provide the first evidence that an Type IIIA - zinc finger protein, Ca1244, plays a crucial role in the activation of the pathogen defense response in plants.

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Development of latent fingerprints contaminated with ethanol on paper surfaces

  • Park, Eun-Jung;Hong, Sungwook
    • Analytical Science and Technology
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    • v.32 no.3
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    • pp.105-112
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    • 2019
  • Fingerprints may be contaminated with ethanol solutions. In order to solve the case, the law enforcement agency may need to visualize the fingerprint from these samples, but the development method has not been studied. The paper with latent fingerprint was contaminated with ethanol solution and then the blurring of ridge detail was observed. As a result, when the copy paper was contaminated with ethanol solutions of less than 75 % (v/v), the amino acid components of latent fingerprint residue blurred but lipid components of latent fingerprint residue didn't blurred. On the other hand, when the paper was contaminated with ethanol solution of more than 80 % (v/v), the amino acid components of latent fingerprint didn't blurred but the lipid components of latent fingerprint blurred. Therefore, it is found that the paper contaminated with ethanol solutions of less than 75 % (v/v) should be treated by oil red O (ORO) enhancing lipid components, and the paper contaminated with ethanol solutions of 80 % (v/v) or more should be treated by 1,2-indandione/zinc (1,2-IND/Zn) enhancing amino acid components. The blurring of ridge detail was not observed when the fingerprints were deposited with fingers contaminated with ethanol solution. This fingerprints were treated with 1,2-IND/Zn or ORO to compare the latent fingerprint development ability, and using 1,2-IND/Zn was able to visualize the latent fingerprint more clearly than using ORO.

Identification of a Novel Human Zinc Finger Gene, ZNF438, with Transcription Inhibition Activity

  • Zhong, Zhaomin;Wan, Bo;Qiu, Yun;Ni, Jun;Tang, Wenwen;Chen, Xinya;Yang, Yun;Shen, Suqin;Wang, Ying;Bai, Meirong;Lang, Qingyu;Yu, Long
    • BMB Reports
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    • v.40 no.4
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    • pp.517-524
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    • 2007
  • There were many different families of zinc finger proteins that contained multiple cysteine and/or histidine residues and used zinc to stabilize their folds. The classical C2H2 zinc finger proteins were the founding members of this superfamily and were among the most abundant proteins in eukaryotic genomes. C2H2 proteins typically contained several C2H2 fingers that made tandem contacts along the DNA. Here we reported a novel C2H2 type zinc finger gene, ZNF438, which encoded 828 amino acids that formed five zinc finger domains. Bioinformatics analysis revealed that the ZNF438 was mapped to human chromosome 10p11.2 and shared 62% identity with rat and mouse homologues. RT-PCR analysis indicated that it was ubiquitously expressed in 18 human adult tissues. With immunofluorescence assay, it was shown that the exogenous Flag-tagged ZNF438 was located in nucleus of COS-7 cells. To further explore the function of ZNF438, we examined the transcriptional activity of ZNF438 protein by transfecting recombinant pM-ZNF438 into mammalian cells. The subsequent analysis based on the duel luciferase assay system showed that ZNF438 was a transcriptional repressor.

Prevalence Rate of Lead Related Subjective Symptoms in Lead Workers (연취급 근로자의 연폭로 수준에 따른 주관적 자각증상 호소율)

  • Jeong, Du-Shin;Kim, Hwa-Sung;Ahn, Kyu-Dong;Lee, Byung-Kook
    • Journal of Preventive Medicine and Public Health
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    • v.26 no.2 s.42
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    • pp.251-267
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    • 1993
  • The relationship between lead related subject symptoms and lead exposure indices was studied in 435 male lead workers in thirteen lead using industries. 212 male office workers who were not exposed to lead occupationally were also studied as a control group. Fourteen lead related symptoms were selected. They were further subdivied into 4 sub-symptom groups such as 1) gastrointestinal, 2) neuromuscular and joint 3) constitutional, and 4) psychological symptoms. Symptom questionnaires were provided to the workers and filled up by themselves and reconfirmed by interviewer(doctor). The test used fer the evaluation of lead exposure were blood lead(PbB), zinc protoporphyrin in whole blood(ZPP), hemoglobin(Hb), hematocrit (Hct), delta-aminolevulinic acid in urine(DALA). The results obtained were as follows; 1. The higher prevalence rate in the sub-group of neuromuscular and joint symptoms was observed in occupationally lead exposed subjects than non-exposed subjects. Among the sub-groups, the most frequent symptom was 'numbness of finger, hands or feet', and the prevalence of the symptom of 'arthralgia', 'weakness of fingers, hands or feet' and 'myalgia' were higher in order. 2. While the symptom which showed the biggest difference of prevalence rate among the 14 symptoms between exposed and non-exposed subjects was 'numbness of fingers, hands or feet', the symptom which showed the highest prevalence rate was 'feeling tired generally' in exposed and non-exposed subjects, but no statistical difference of symptom prevalence were observed. 3. In total study population, PbB and ZPP had dose-response relationship with 4 symtoms of neuromuscular and joint symptoms ('numbness of finger, hands or feet', 'arthralgia', 'weakness of fingers, hands or feet' and 'myalgia') and one symptom of gastrointestinal group('intermittent pains in lower abdomen'). 4. In lead exposed workers, only neuromuscular and joint symptoms group showed dose-response relationship with PbB and ZPP, 5. In lead exposed workers, the prevalance rate of overall symptoms of lead workers with age below 39 years was higher than that of lead workers with age above 40. While neuromuscular and joint symptoms group had a dose-response relationship with PbB in former group, it had a dose-response relationship with ZPP in latter group. 6. Age adjusted odds ratios of symptoms of non-exposed with exposed and odds ratios of low exposed with high exposed workers showed the dose-response relationship of lead exposure with neuromuscular and joint symptoms group('numbness of fingers, hands or feet', 'arthralgia', 'weakness of fingers, hands or feet' and 'myalgia') and gastrointestinal symptoms group('intermittent pains in lower abdoman').

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