• Title/Summary/Keyword: i-motif

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Cloning and molecular characterization of a new fungal xylanase gene from Sclerotinia sclerotiorum S2

  • Ellouze, Olfa Elleuch;Loukil, Sana;Marzouki, Mohamed Nejib
    • BMB Reports
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    • v.44 no.10
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    • pp.653-658
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    • 2011
  • Sclerotinia sclerotiorum fungus has three endoxylanases induced by wheat bran. In the first part, a partial xylanase sequence gene (90 bp) was isolated by PCR corresponding to catalytic domains (${\beta}5$ and ${\beta}6$ strands of this protein). The high homology of this sequence with xylanase of Botryotinia fuckeliana has permitted in the second part to amplify the XYN1 gene. Sequence analysis of DNA and cDNA revealed an ORF of 746 bp interrupted by a 65 bp intron, thus encoding a predicted protein of 226 amino acids. The mature enzyme (20.06 kDa), is coded by 188 amino acid (pI 9.26). XYN1 belongs to G/11 glycosyl hydrolases family with a conserved catalytic domain containing $E_{86}$ and $E_{178}$ residues. Bioinformatics analysis revealed that there was no Asn-X-Ser/Thr motif required for N-linked glycosylation in the deduced sequence however, five O-glycosylation sites could intervene in the different folding of xylanses isoforms and in their secretary pathway.

Investigation of functional roles of transcription termination factor-1 (TTF-I) in HIV-1 replication

  • Park, Seong-Hyun;Yu, Kyung-Lee;Jung, Yu-Mi;Lee, Seong-Deok;Kim, Min-Jeong;You, Ji-Chang
    • BMB Reports
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    • v.51 no.7
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    • pp.338-343
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    • 2018
  • Transcription termination factor-1 (TTF-I) is an RNA polymerase 1-mediated transcription terminator and consisting of a C-terminal DNA-binding domain, central domain, and N-terminal regulatory domain. This protein binds to a so-called 'Sal box' composed of an 11-base pair motif. The interaction of TTF-I with the 'Sal box' is important for many cellular events, including efficient termination of RNA polymerase-1 activity involved in pre-rRNA synthesis and formation of a chromatin loop. To further understand the role of TTF-I in human immunodeficiency virus (HIV)-I virus production, we generated various TTF-I mutant forms. Through a series of studies of the over-expression of TTF-I and its derivatives along with co-transfection with either proviral DNA or HIV-I long terminal repeat (LTR)-driven reporter vectors, we determined that wild-type TTF-I downregulates HIV-I LTR activity and virus production, while the TTF-I Myb-like domain alone upregulated virus production, suggesting that wild-type TTF-I inhibits virus production and trans-activation of the LTR sequence; the Myb-like domain of TTF-I increased virus production and trans-activated LTR activity.

Regulation of toll-like receptors expression in muscle cells by exercise-induced stress

  • Park, Jeong-Woong;Kim, Kyung-Hwan;Choi, Joong-Kook;Park, Tae Sub;Song, Ki-Duk;Cho, Byung-Wook
    • Animal Bioscience
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    • v.34 no.10
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    • pp.1590-1599
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    • 2021
  • Objective: This study investigates the expression patterns of toll-like receptors (TLRs) and intracellular mediators in horse muscle cells after exercise, and the relationship between TLRS expression in stressed horse muscle cells and immune cell migration toward them. Methods: The expression patterns of the TLRs (TLR2, TLR4, and TLR8) and downstream signaling pathway-related genes (myeloid differentiation primary response 88 [MYD88]; activating transcription factor 3 [ATF3]) are examined in horse tissues, and horse peripheral blood mononuclear cells (PBMCs), polymorphonuclear cells (PMNs) and muscles in response to exercise, using the quantitative reverse transcription-polymerase chain reaction (qPCR). Expressions of chemokine receptor genes, i.e., C-X-C motif chemokine receptor 2 (CXCR2) and C-C motif chemokine receptor 5 (CCR5), are studied in PBMCs and PMNs. A horse muscle cell line is developed by transfecting SV-T antigen into fetal muscle cells, followed by examination of muscle-specific genes. Horse muscle cells are treated with stressors, i.e., cortisol, hydrogen peroxide (H2O2), and heat, to mimic stress conditions in vitro, and the expression of TLR4 and TLR8 are examined in stressed muscle cells, in addition to migration activity of PBMCs toward stressed muscle cells. Results: The qPCR revealed that TLR4 message was expressed in cerebrum, cerebellum, thymus, lung, liver, kidney, and muscle, whereas TLR8 expressed in thymus, lung, and kidney, while TLR2 expressed in thymus, lung, and kidney. Expressions of TLRs, i.e., TLR4 and TLR8, and mediators, i.e., MYD88 and ATF3, were upregulated in muscle, PBMCs and PMNs in response to exercise. Expressions of CXCR2 and CCR5 were also upregulated in PBMCs and PMNs after exercise. In the muscle cell line, TLR4 and TLR8 expressions were upregulated when cells were treated with stressors such as cortisol, H2O2, and heat. Migration of PBMCs toward stressed muscle cells was increased by exercise and oxidative stresses, and combinations of these. Treatment with methylsulfonylmethane (MSM), an antioxidant on stressed muscle cells, reduced migration of PBMCs toward stressed muscle cells. Conclusion: In this study, we have successfully cultured horse skeletal muscle cells, isolated horse PBMCs, and established an in vitro system for studying stress-related gene expressions and function. Expression of TLR4, TLR8, CXCR2, and CCR5 in horse muscle cells was higher in response to stressors such as cortisol, H2O2, and heat, or combinations of these. In addition, migration of PBMCs toward muscle cells was increased when muscle cells were under stress, but inhibition of reactive oxygen species by MSM modulated migratory activity of PBMCs to stressed muscle cells. Further study is necessary to investigate the biological function(s) of the TLR gene family in horse muscle cells.

Characterization of αX I-Domain Binding to Receptors for Advanced Glycation End Products (RAGE)

  • Buyannemekh, Dolgorsuren;Nham, Sang-Uk
    • Molecules and Cells
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    • v.40 no.5
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    • pp.355-362
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    • 2017
  • The ${\beta}2$ integrins are cell surface transmembrane proteins regulating leukocyte functions, such as adhesion and migration. Two members of ${\beta}2$ integrin, ${\alpha}M{\beta}2$ and ${\alpha}X{\beta}2$, share the leukocyte distribution profile and integrin ${\alpha}X{\beta}2$ is involved in antigen presentation in dendritic cells and transendothelial migration of monocytes and macrophages to atherosclerotic lesions. ${\underline{R}}eceptor$ for ${\underline{a}}dvanced$ ${\underline{g}}lycation$ ${\underline{e}}nd$ ${\underline{p}}roducts$ (RAGE), a member of cell adhesion molecules, plays an important role in chronic inflammation and atherosclerosis. Although RAGE and ${\alpha}X{\beta}2$ play an important role in inflammatory response and the pathogenesis of atherosclerosis, the nature of their interaction and structure involved in the binding remain poorly defined. In this study, using I-domain as a ligand binding motif of ${\alpha}X{\beta}2$, we characterize the binding nature and the interacting moieties of ${\alpha}X$ I-domain and RAGE. Their binding requires divalent cations ($Mg^{2+}$ and $Mn^{2+}$) and shows an affinity on the sub-micro molar level: the dissociation constant of ${\alpha}X$ I-domains binding to RAGE being $0.49{\mu}M$. Furthermore, the ${\alpha}X$ I-domains recognize the V-domain, but not the C1 and C2-domains of RAGE. The acidic amino acid substitutions on the ligand binding site of ${\alpha}X$ I-domain significantly reduce the I-domain binding activity to soluble RAGE and the alanine substitutions of basic amino acids on the flat surface of the V-domain prevent the V-domain binding to ${\alpha}X$ I-domain. In conclusion, the main mechanism of ${\alpha}X$ I-domain binding to RAGE is a charge interaction, in which the acidic moieties of ${\alpha}X$ I-domains, including E244, and D249, recognize the basic residues on the RAGE V-domain encompassing K39, K43, K44, R104, and K107.

Recovery of TRIM25-Mediated RIG-I Ubiquitination through Suppression of NS1 by RNA Aptamers

  • Woo, Hye-Min;Lee, Jin-Moo;Kim, Chul-Joong;Lee, Jong-Soo;Jeong, Yong-Joo
    • Molecules and Cells
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    • v.42 no.10
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    • pp.721-728
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    • 2019
  • Non-structural protein 1 (NS1) of influenza virus has been shown to inhibit the innate immune response by blocking the induction of interferon (IFN). In this study, we isolated two single-stranded RNA aptamers specific to NS1 with $K_d$ values of $1.62{\pm}0.30nM$ and $1.97{\pm}0.27nM$, respectively, using a systematic evolution of ligand by exponential enrichment (SELEX) procedure. The selected aptamers were able to inhibit the interaction of NS1 with tripartite motif-containing protein 25 (TRIM25), and suppression of NS1 enabled retinoic acid inducible gene I (RIG-I) to be ubiquitinated regularly by TRIM25. Additional luciferase reporter assay and quantitative real-time PCR (RT-PCR) experiments demonstrated that suppression of NS1 by the selected aptamers induced IFN production. It is noted that viral replication was also inhibited through IFN induction in the presence of the selected aptamers. These results suggest that the isolated aptamers are strongly expected to be new therapeutic agents against influenza infection.

A Study on the Wedding Dress Design of Korea Images - Focused on Pear Blossoms Pattern and Dang-$\breve{u}$i(唐衣) Images - (한국적 이미지의 웨딩드레스 디자인 연구 - 이화문(梨花紋)과 당의(唐衣)이미지를 중심으로 -)

  • Lee, Min-Jung;Cho, Kyu-Hwa
    • Journal of Fashion Business
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    • v.13 no.5
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    • pp.23-36
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    • 2009
  • The purpose of this research is to design and to produce actual high value added wedding dress comprised of the pear blossoms pattern(梨花紋), which is one of the traditional patterns which was not researched to this day, and of the Dang-$\breve{u}$i(唐衣) image which reflects our concept of beauty as if as a means of conducting empirical study for the application of traditional Korean patterns to the contemporary application. pear blossoms effectively signifie the image of beautiful bride thanks to the pure, graceful and clean image of white flower, both in the external and internal aspects. The Dang-$\breve{u}$i of the Chosun Dynasty, which is another motif is one of the representative clothing that expresses the beauty of the curve when it comes to the Korean culture as traditional wedding dress. Attempt was made to develop design for wedding dress that expresses the Korean style beauty that combined the tradition and the modern. Towards this end, the project was composed of the Korean-like Ewha, image of the Dang-$\breve{u}$i I, II and III to design three dresses. The Dress I presents the image of pure, elegant and beautiful bride, while Dress II represents the pure and city like trendiness of the Korean women today as bride. Dress III was made centered on the image of elegant, and chaste bride. As for the materials used, hand-woven silk and silk organza used for the traditional clothes. As for the ornaments, embroidery, quilted work, beads, corsage, burn-out and other techniques were used for expression.

The β Subunit of Heterotrimeric G Protein Interacts Directly with Kinesin Heavy Chains, Kinesin-I (Kinesin-I의 kinesin heavy chains과 직접 결합하는 heterotrimeric G protein의 β subunit의 규명)

  • Seog, Dae-Hyun
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1166-1172
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    • 2010
  • Kinesin-I exists as a tetramer of two heavy chains (KHCs, also called KIF5s), which contain the amino (N)-terminal motor domain and carboxyl (C)-terminal domain, as well as two light chains (KLCs), which bind to the KIF5s (KIF5A, KIF5B and KIF5C) stalk region. To identify the interaction proteins for KIF5A, yeast two-hybrid screening was performed and a specific interaction with the ${\beta}$ subunit of heterotrimeric G proteins ($G{\beta}$) was found. $G{\beta}$ bound to the amino acid residues between 808 and 935 of KIF5A and to other KIF5 members in the yeast two-hybrid assay. The WD40 repeat motif of $G{\beta}$ was essential for interaction with KIF5A. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KIF5s specifically co-immunoprecipitated KIF5s associated with heterotrimeric G proteins from mouse brain extracts. These results suggest that kinesin-I motor protein transports heteroterimeric G protein attachment vesicles along microtubules in the cell.

Purification and Characterization of Repressor of Temperate S. aureus Phage Φ11

  • Das, Malabika;Ganguly, Tridib;Chattoraj, Partho;Chanda, Palas Kumar;Bandhu, Amitava;Lee, Chia Yen;Sau, Subrata
    • BMB Reports
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    • v.40 no.5
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    • pp.740-748
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    • 2007
  • To gain insight into the structure and function of repressor proteins of bacteriophages of gram-positive bacteria, repressor of temperate Staphylococcus aureus phage ${\phi}11$ was undertaken as a model system here and purified as an N-terminal histidine-tagged variant (His-CI) by affinity chromatography. A ~19 kDa protein copurified with intact His-CI (~ 30 kDa) at low level was resulted most possibly due to partial cleavage at its Ala-Gly site. At ~10 nM and higher concentrations, His-CI forms significant amount of dimers in solution. There are two repressor binding sites in ${\phi}11$ cI-cro intergenic region and binding to two sites occurs possibly by a cooperative manner. Two sites dissected by HincII digestion were designated operators $O_L$ and $O_R$, respectively. Equilibrium binding studies indicate that His-CI binds to $O_R$ with a little more strongly than $O_L$ and binding species is probably dimeric in nature. Interestingly His-CI binding affinity reduces drastically at elevated temperatures ($32-42^{\circ}C$). Both $O_L$ and $O_R$ harbor a nearly identical inverted repeat and studies show that ${\phi}11$ repressor binds to each repeat efficiently. Additional analyses indicate that ${\phi}11$ repressor, like $\lambda$ repressor, harbors an N-terminal domain and a C-terminal domain which are separated by a hinge region. Secondary structure of ${\phi}11$ CI even nearly resembles to that of $\lambda$ phage repressor though they differ at sequence level. The putative N-terminal HTH (helix-turn-helix) motif of ${\phi}11$ repressor belongs to the HTH -XRE-family of proteins and shows significant identity to the HTH motifs of some proteins of evolutionary distant organisms but not to HTH motifs of most S. aureus phage repressors.

Analysis of the solution structure of the human antibiotic peptide dermcidin and its interaction with phospholipid vesicles

  • Jung, Hyun-Ho;Yang, Sung-Tae;Sim, Ji-Yeong;Lee, Seung-Kyu;Lee, Ju-Yeon;Kim, Ha-Hyung;Shin, Song-Yub;Kim, Jae-Il
    • BMB Reports
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    • v.43 no.5
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    • pp.362-368
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    • 2010
  • Dermcidin is a human antibiotic peptide that is secreted by the sweat glands and has no homology to other known antimicrobial peptides. As an initial step toward understanding dermcidin's mode of action at bacterial membranes, we used homonuclear and heteronuclear NMR to determine the conformation of the peptide in 50% trifluoroethanol solution. We found that dermcidin adopts a flexible amphipathic $\alpha$-helical structure with a helix-hinge-helix motif, which is a common molecular fold among antimicrobial peptides. Spin-down assays of dermcidin and several related peptides revealed that the affinity with which dermcidin binds to bacterial-mimetic membranes is primarily dependent on its amphipathic $\alpha$-helical structure and its length (>30 residues); its negative net charge and acidic pI have little effect on binding. These findings suggest that the mode of action of dermcidin is similar to that of other membrane-targeting antimicrobial peptides, though the details of its antimicrobial action remain to be determined.

A Study Conceming the Designs of the Afro Fashion (아프로 패션에 관(關)한 디자인 연구(硏究))

  • Kim, Chi-Eun;Cho, Kyu-Hwa
    • Journal of Fashion Business
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    • v.2 no.1
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    • pp.25-34
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    • 1998
  • This study has it's purpose in examining the materialized background. characteristic of African traditional costume and the A fro fashion of the year 1960. and it's influence on the contemporary fashion. Then the findings are applied to suggest a new way of image creation. 1960' s was the year in which people tried to free oneself from the ruling culture of the social standards. war. and the development of science. By such movement. people started to get interested in the environment and ecology. This then lead to the interest of the rights of the minority. With the youngs as the central figure A fro hair style and dashiki appeared as the street fashion. The characteristics of African costume applied to A fro fashion in 1990' s is as below. First. the North African style. Djellaba. and wrap style in the most common silhouette. Second. heavy materials such as stone. copper. silver. and gold are used. Necklace can be classified according to it's simple but. modern style. delicate but grand style. Bracelets are however. broad in width and many rings are worn widely. Third. Multi colored stripe and prints inspired by tattoo and deformation using red brown. dark beige. and orange are printed on textiles such as see through. Fourth. image of tattoo and deformation are applied to make-up. A fro hair and corn beads are also African taste. Fifth. African taste in recent fashion showed off the black beauty by appointing black models at the collection. In analyzing the study done above. characteristic images of African costume. accessaries and body painting was applied in presenting 3 creative designs. The first design named "Geometric I", took it's motif from the geometric pattern of the body painting with the afro hair. "Geometric II", the second design, showed it's application of geometric pattern of mutilation and the silhouette of the costume by using the see through. The third design called the "Geometric III" showed that the aesthetic and decorative side of clothing can be satisfied by applying various form and color of accessaries as the motif. A fro fashion is chosen as one of the folklore mood and it's beauty is conveyed on till today.

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