• Title/Summary/Keyword: DNA interaction

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Differential expression and in situ localization of a pepper defensin (CADEFl) gene in response to pathogen infection, abiotic elicitors and environmental stresses in Capsium annuum

  • Do, Hyun-Mee;Lee, Sung-Chul;Jung, Ho-Won;Hwang, Byung-Kook
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.78.2-79
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    • 2003
  • Pepper defensin ( CADEFl) clone was isolated from cDNA library constructed from pepper leaves infected with avirulent strain Bv5-4a of Xanthomonu campestris pv. vesicatoria. The deduced amino acid sequence of CADEFl is 82-64% identical to that of other plant defensins. Putative protein encoded by CADEFl gene consists of 78 amino acids and 8 conserved cysteine residues to form four structure-stabilizing disulfide bridges. Transcription of the CADEF1 gene was earlier and stronger induced by X campestris pv. vesicatoria infection in the incompatible than in the compatible interaction. CADEF1 mRNA was constitutively expressed in stem, root and green fruit of pepper. Transcripts of CADEFl gene drastically accumulated in pepper leaf tissues treated With Salicylic acid (SA), methyl jasmonate (MeJA), abscisic acid (ABA), hydrogen Peroxide (H$_2$O$_2$), benzothiadiazole (BTH) and DL-${\beta}$-amino-n-butyric acid (BABA). In situ hybridization results revealed that CADEF1 mRNA was localized in the phloem areas of vascular bundles in leaf tissues treated with exogenous SA, MeJA and ABA. Strong accumulation of CADEF1 mRNA occurred in pepper leaves in response to wounding, high salinity and drought stress. These results suggest that bacterial pathogen infection, abiotic elicitors and some environmental stresses may play a significant role in signal transduction pathway for CADEF1 gene expression.

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Isolation and Characterization of Pathogen inducible Leucine Zipper containing Gene from rice (Oryza sativa L. cv. Dongjin)

  • Park, Sang-Ryeol;Song, Hae-Sook;Moon, Kyung-Mi;Hwang, Duk-Ju;Kim, Tae-Ho;Han, Seong-Sook;Go, Seung-Joo;Byun, Myung-Ok
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.83.2-83
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    • 2003
  • A full length cDNA, OsLEUZIP, encoding leucine zipper containing protein from rice EST of rice (0ryza sativa L. cv. Dongjin) treated Xanthomonas oryzae pv. oryzae 10331. OsLEUZIP contains 1,227 bp nucleotides and encodes a protein of 408 amino acid residues with predicted molecular weight of 47,229 Da. The deduced amino acid sequence of OsLEUZIP has consensus sequence of leucine zipper from PROSITE (PDOC00029), L-X(6)-L-X(6)-L-X(6) -L. OsLEUZIP gene were preferentially induced in rice during incompatible interaction with Xanthomonas oryzae pv. oryzae 10331 and Pyracuraria grisea KJ-301. Expression of OsLEUZIP gene was also induced by treatment of abiotics such as ethephon and ABA. Our data represented in this study suggesting that OsLEUZIP gene may play an important role in the rice defense-related. Further studies of this gene, overexpression in rice, yeast-two hybrid assay, electrophoretic mobility shift assay and northern blot analyses of transgenic plant, would be useful to elucidate the role of the OsLEUZIP gene in defense responses of rice.

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CaGe: A Web-Based Cancer Gene Annotation System for Cancer Genomics

  • Park, Young-Kyu;Kang, Tae-Wook;Baek, Su-Jin;Kim, Kwon-Il;Kim, Seon-Young;Lee, Do-Heon;Kim, Yong-Sung
    • Genomics & Informatics
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    • v.10 no.1
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    • pp.33-39
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    • 2012
  • High-throughput genomic technologies (HGTs), including next-generation DNA sequencing (NGS), microarray, and serial analysis of gene expression (SAGE), have become effective experimental tools for cancer genomics to identify cancer-associated somatic genomic alterations and genes. The main hurdle in cancer genomics is to identify the real causative mutations or genes out of many candidates from an HGT-based cancer genomic analysis. One useful approach is to refer to known cancer genes and associated information. The list of known cancer genes can be used to determine candidates of cancer driver mutations, while cancer gene-related information, including gene expression, protein-protein interaction, and pathways, can be useful for scoring novel candidates. Some cancer gene or mutation databases exist for this purpose, but few specialized tools exist for an automated analysis of a long gene list from an HGT-based cancer genomic analysis. This report presents a new web-accessible bioinformatic tool, called CaGe, a cancer genome annotation system for the assessment of candidates of cancer genes from HGT-based cancer genomics. The tool provides users with information on cancer-related genes, mutations, pathways, and associated annotations through annotation and browsing functions. With this tool, researchers can classify their candidate genes from cancer genome studies into either previously reported or novel categories of cancer genes and gain insight into underlying carcinogenic mechanisms through a pathway analysis. We show the usefulness of CaGe by assessing its performance in annotating somatic mutations from a published small cell lung cancer study.

Telomere association of Oryza sativa telomere repeat-binding factor like 1 and its roles in telomere maintenance and development in rice, Oryza sativa L.

  • Byun, Mi Young;Cui, Li Hua;Lee, Hyoungseok;Kim, Woo Taek
    • BMB Reports
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    • v.51 no.11
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    • pp.578-583
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    • 2018
  • Telomeres are specialized nucleoprotein complexes that function to protect eukaryotic chromosomes from recombination and erosion. Several telomere binding proteins (TBPs) have been characterized in higher plants, but their detailed in vivo functions at the plant level are largely unknown. In this study, we identified and characterized OsTRFL1 (Oryza sativa Telomere Repeat-binding Factor Like 1) in rice, a monocot model crop. Although OsTRFL1 did not directly bind to telomere repeats $(TTTAGGG){_4}$ in vitro, it was associated with telomeric sequences in planta. OsTRFL1 interacted with rice TBPs, such as OsTRBF1 and RTBP1, in yeast and plant cells as well as in vitro. Thus, it seems likely that the association of OsTRFL1 with other TBPs enables OsTRFL1 to bind to telomeres indirectly. T-DNA inserted OsTRFL1 knock-out mutant rice plants displayed significantly longer telomeres (6-25 kb) than those (5-12 kb) in wild-type plants, indicating that OsTRFL1 is a negative factor for telomere lengthening. The reduced levels of OsTRFL1 caused serious developmental defects in both vegetative and reproductive organs of rice plants. These results suggest that OsTRFL1 is an essential factor for the proper maintenance of telomeres and normal development of rice.

Effects of FIS Protein on rnpB Transcription in Escherichia coli

  • Choi, Hyun-Sook;Kim, Kwang-sun;Park, Jeong Won;Jung, Young Hwan;Lee, Younghoon
    • Molecules and Cells
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    • v.19 no.2
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    • pp.239-245
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    • 2005
  • Factor for inversion stimulation (FIS), the Escherichia coli protein, is a positive regulator of the transcription of genes that encode stable RNA species, such as rRNA and tRNA. Transcription of the rnpB gene encoding M1 RNA, the catalytic subunit of E. coli RNase P, rapidly declines under stringent conditions, as does that of other stable RNAs. There are multiple putative FIS binding sites upstream of the rnpB promoter. We tested whether FIS binds to these sites, and if so, how it affects rnpB transcription. In vitro binding assays revealed specific binding of FIS to multiple sites in the rnpB promoter region. Interestingly, FIS bound not only to the upstream region of the promoter, but also to the region from +4 to +18. FIS activated rnpB transcription in vitro, but the level of activation was much lower than that of the rrnB promoter for rRNA. We also examined the effects of FIS on rnpB transcription in vivo using isogenic $fis^+$ and $fis^-$ strains. rnpB transcription was higher in the $fis^-$ than the $fis^+$ cells during the transitions from lag to exponential phase, and from exponential to stationary phase.

Functional Gene Analysis to Identify Potential Markers Induced by Benzene in Two Different Cell Lines, HepG2 and HL-60

  • Kim, Youn-Jung;Song, Mi-Kyung;Sarma, Sailendra Nath;Choi, Han-Saem;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.4 no.3
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    • pp.183-191
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    • 2008
  • Volatile organic compounds (VOCs) are common constituents of cleaning and degreasing agents, paints, pesticides, personal care products, gasoline and solvents. And VOCs are evaporated at room temperature and most of them exhibit acute and chronic toxicity to human. Benzene is the most widely used prototypical VOC and the toxic mechanisms of them are still unclear. The multi-step process of toxic mechanism can be more fully understood by characterizing gene expression changes induced in cells by toxicants. In this study, DNA microarray was used to monitor the expression levels of genes in HepG2 cells and HL-60 cells exposed to the benzene on IC20 and IC50 dose respectively. In the clustering analysis of gene expression profiles, although clusters of HepG2 and HL-60 cells by benzene were divided differently, expression pattern of many genes observed similarly. We identified 916 up-regulated genes and 1,144 down-regulated genes in HepG2 cells and also 1,002 up-regulated genes and 919 down-regulated genes in HL-60 cells. The gene ontology analysis on genes expressed by benzene in HepG2 and HL-60 cells, respectively, was performed. Thus, we found some principal pathways, such as, focal adhesion, gap junction and signaling pathway in HepG2 cells and toll-like receptor signaling pathway, MAPK signaling pathway, p53 signaling pathway and neuroactive ligand-receptor interaction in HL-60 cells. And we also found 16 up-regulated and 14 down-regulated commonly expressed total 30 genes that belong in the same biological process like inflammatory response, cell cycle arrest, cell migration, transmission of nerve impulse and cell motility in two cell lines. In conclusion, we suggest that this study is meaningful because these genes regarded as strong potential biomarkers of benzene independent of cell type.

Cell Surface Interaction with Expression of Fas Ligand Mediates Prolactin-Induced Apoptosis In Rat Luteal Cell Culture (Rat 황체세포 배양에 있어서 Prolactin에 의한 황체퇴행 및 Fas Ligand의 발현)

  • 장규태;박미령;선동수;윤창현
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.179-190
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    • 1998
  • Prolactin (PRL) surge in cycling rats at proestrous afternoon has previously been reported as an inducer of apoptotic cell death of luteal cells. This death-inducing action of PRL seeins unusual, because PRL can he categorized as a cell-survival factor, if other known physiological functions of PRL are taken into account. In this study, the apoptotic action of PRL was assessed in cultured cells prepared from rat luteal tissue and underlying molecular /cellular mechanism of PRL-induced luteolysis was analyzed. The latest crop of corpora lutea (CLs) were enucleated from rat ovaries at 18:00 h on the proestrous day before the next ovulation. Donor rats were pretreated with CB154, a dopamine agonist, in order to he exempted from the endogenous PRL surge. The harvested GLs were dispersed and cultured with or without PRL (2$\mu$g /ml) for 24 or 48 h. An addition of PRL to the culture medium changed the parameters indicative of cell death via apoptosis: a decrease in cell viability (MTT) and an increase in chromatin condensation. Most of the DNA breakdown in nuclei induced by PRL occurred in steroidogenic cells which were identified by 3$\beta$-HSD activity staining, and the number of 3$\beta$-HSD-positivecells were significantly decreased. Interestingly, most of the cells with an apoptotic nucleus adhered to one or more intact and seemingly non-steroidogenic cells. Because the expression of Fas has heen shown to be abundant in murine ovary, and Fas is known to have an exact physiological role in occurrence of apoptotic cell death, the membrane form-Fas ligand (rnFasL) was quantified in the cell lysate. An addition of PRL increased expression of mFasL. Moreover, an addition of concanavalin A (ConA), a T-cell specific activator, in place of PRL, enhanced the apoptotic parameters. Cumulatively, the apoptotic PRL action was addressed to cells unknown than steroidogenic lute~ cells. The most prohable candidate for the direct target cells is Tcells in the luteal tissue that can express mFasL in response to PRL.

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Modulation of Cytotoxicity by Nitric Oxide Donors during Treatment of Glioma with Anticancer Drugs

  • Park, Jeong-Jae;Kang, Jong-Sool;Lee, Hyun-Sung;Lee, Jong-Soo;Lee, Young-Ha;Youm, Jin-Young
    • Journal of Korean Neurosurgical Society
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    • v.38 no.5
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    • pp.366-374
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    • 2005
  • Objective : Nitric oxide[NO] is implicated in a wide range of biological processes in tumors and is produced in glioma. To investigate the role of NO and its interaction with the tumoricidal effects of anticancer drugs, we study the antitumor activities of NO donors, with or without anticancer drugs, in human glioma cell lines. Methods : U87MG and U373MG cells were treated with the NO donors sodium nitroprusside[SNP] and S-nitroso-N-acetylpenicillamine[SNAP], alone or in combination with the anticancer drugs 1,3-bis[2-chloroethyl]-1-nitrosourea[BCNU] and cisplatin. Cell viability, cell proliferation, DNA fragmentation, nitrite level, and the expression of Bcl-2 and Bax were determined. Results : NO was markedly increased after treatment with SNP or SNAP; however, the addition of the anticancer drugs did not significantly affect NO production NO donors or anticancer drugs reduced glioma cell viability and, in combination, acted synergistically to further decrease cell viability in a dose- and time-dependent manner. Cell proliferation was inhibited and apoptosis were enhanced by combined treatment. Bax expression was increased by combined treatment, whereas Bcl-2 expression was reduced. The antitumor cytotoxicity of NO donors and anticancer drugs differed according to cell type. Conclusion : BCNU or cisplatin can inhibit cell viability and proliferation of glioma cells and can induce apoptosis. These effects are further enhanced by the addition of a NO donor which modulates the antitumor cytotoxicity of chemotherapy depending on cell type. Further biological, chemical, and toxicological studies of NO are required to clarify its mechanism of action in glioma.

Molecular determinants of the host specificity by Xanthomonas spp.

  • Heu, Sunggi;Choi, Min-Seon;Park, Hyoung-Joon;Lee, Seung-Don;Ra, Dong-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2004.10a
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    • pp.65-67
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    • 2004
  • During initial interactions of bacteria with their host plants, most plants recognize the bacterial infections and repel the pathogen by plant defense mechanism. The most active plant defense mechanism is the hypersensitive response (HR) which is the localized induced cell death in the plant at the site of infection by a pathogen. A primary locus induced in gram-negative phytopathogenic bacteria during this initial interaction is the Hrp locus. The Hrp locus is composed of a cluster of genes that encodes the bacteral Type 111 machinery that is involved in the secretion and translocation of effector proteins to the plant cell. DNA sequence analysis of hrp gene in phytopathogenic bacteria has revealed a Hrp pathogenicity is]and (PAI) with a tripartite mosaic structure. For many gram-negative pathogenic bacteria, colonization of the host's tissue depends on the type III protein secretion system (TTSS) which secrets and translocates effector proteins into the host cell. Effectors can be divided into several groups including broad host range effectors, host specific effectors, disease specific effectors, and effectors inhibit host defenses. The role of effectors carrying LRR domain in plant resistance is very elusive since most known plant resistance gene carry LRR domain. Host specific effectors such as several avr gene products are involved in the determination of the host specificity. Almost all the phytopathogenic Xanthomonas spp. carry avrBs1, avrBs2, and avrBs3 homologs. Some strains of X. oryzae pv. oryzae carry more than 10 copies of avrBs3 homologs. However, the functions of all those avr genes in host specificity are not characterized well.;

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Differential Expression of Gene Profiles in MRGX-treated Lung Cancer

  • Kwon, Yong-Kyun;Lee, Seung-Yeul;Kang, Hwan-Soo;Sung, Jung-Suk;Cho, Chong-Kwan;Yoo, Hwa-Seung;Shin, Seungjin;Choi, Jong-Soon;Lee, Yeon-Weol;Jang, Ik-Soon
    • Journal of Pharmacopuncture
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    • v.16 no.3
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    • pp.30-38
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    • 2013
  • Objectives: Modified regular ginseng extract (MRGX) has stronger anti-cancer activity-possessing gensenoside profiles. Methods: To investigate changes in gene expression in the MRGX-treated lung cancer cells (A549), we examined genomic data with cDNA microarray results. After completing the gene-ontology-based analysis, we grouped the genes into up-and down-regulated profiles and into ontology-related regulated genes and proteins through their interaction network. Results: One hundred nine proteins that were up- and down-regulated by MRGX were queried by using IPA. IL8, MMP7 and PLAUR and were found to play a major role in the anti-cancer activity in MRGX-treated lung cancer cells. These results were validated using a Western blot analysis and a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis. Conclusions: Most MRGX-responsive genes are up-regulated transiently in A549 cells, but down-regulated in a sustained manner in lung cancer cells.