• Title/Summary/Keyword: protein tissues

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Cancer-Upregulated Gene 2 (CUG2), a New Component of Centromere Complex, Is Required for Kinetochore Function

  • Kim, Hyejin;Lee, Miae;Lee, Sunhee;Park, Byoungwoo;Koh, Wansoo;Lee, Dong Jun;Lim, Dae-Sik;Lee, Soojin
    • Molecules and Cells
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    • v.27 no.6
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    • pp.697-701
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    • 2009
  • We previously identified cancer-upregulated gene 2 (CUG2) as a commonly up-regulated gene in various human cancer tissues, especially in ovary, liver, and lung (Lee et al., 2007a). CUG2 was determined to be a nuclear protein that exhibited high proto-oncogenic activities when overexpressed in NIH3T3 mouse fibroblast cells. To identify other cellular functions of CUG2, we performed yeast two-hybrid screening and identified CENP-T, a component of CENP-A nucleosome complex in the centromere, as an interacting partner of CUG2. Moreover, CENP-A, the principle centromeric determinant, was also found in complex with CENP-T/CUG2. Immunofluorescent staining revealed the co-localization of CUG2 with human centromeric markers. Inhibition of CUG2 expression drastically affected cell viability by inducing aberrant cell division. We propose that CUG2 is a new component of the human centromeric complex that is required for proper chromosome segregation during mitosis.

Association of Clusterin Polymorphisms with Coronary Heart Disease in Koreans

  • Kim, Jun-Young;Kim, Su-Won;Lee, In-Kyu;Yoo, Min
    • Biomedical Science Letters
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    • v.14 no.1
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    • pp.55-58
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    • 2008
  • Clusterin is an 80 kDa heterodimetric glycosylated protein and it plays diverse biological roles in various tissues and organs. Clusterin is reported to be associated with the pathogenesis of coronary artery disease and atherosclerosis. Therefore, we investigated the genotype for the A/G polymorphism at the position 4,183 of clusterin gene in Koreans and compared genotype of patients with control group. 100 patients (Male 63, Female 37), who previously underwent coronary artery stent due to ischemic heart disease and 100 controls (Male 36, Female 64) participated in this study. There was a significant association between 4,183 A/G polymorphism in clusterin gene and coronary artery disease (CAD). The present study shows that clusterin gene A/G polymorphism may be associated with the pathogenesis of CAD. Further studies with larger population may be needed for the development of diagnostic methods at genetic level such as DNA chip.

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The Korean Traditional Medicine Gyeongshingangjeehwan Reduces Lipid Accumulation in Skeletal Muscle and C2C12 Cells

  • Yoon, Mi-Chung
    • Biomedical Science Letters
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    • v.17 no.4
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    • pp.283-289
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    • 2011
  • Our previous study demonstrated that the Korean traditional medicine Gyeongshingangjeehwan (GGEx) activates AMP-activated protein kinase (AMPK) and peroxisome proliferator-activated receptor ${\alpha}$ ($PPAR{\alpha}$) critical for fatty acid oxidation in skeletal muscle and C2C12 skeletal muscle cells. Thus, we examined whether GGEx can reduce lipid accumulation in these cells and tissues. After obese and type 2 diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats were treated with GGEx, we studied the effects of GGEx on skeletal muscle lipid accumulation. The effects of GGEx and/or the AMPK inhibitor compound C on lipid accumulation and expression of AMPK and $PPAR{\alpha}$ were measured in C2C12 skeletal muscle cells. Compared with lean Long-Evans Tokushima Otsuka rats, obese OLETF rats had increased triglyceride droplets. However, administration of GGEx to OLETF rats for 8 weeks significantly decreased triglyceride droplets in skeletal muscle. Consistent with the $in$ $vivo$ data, GGEx inhibited lipid accumulation, the degree of which was comparable to Wy14,643, the potent activator of $PPAR{\alpha}$. GGEx also increased skeletal muscle mRNA levels of AMPK${\alpha}1$, AMPK${\alpha}2$, and $PPAR{\alpha}$. However, compound C inhibited these effects in C2C12 cells. These results suggest that GGEx suppresses skeletal muscle lipid accumulation and this process may be mediated by AMPK and $PPAR{\alpha}$ activation.

Biochemical Changes in Sorghum Leaves Infected with Leaf Spot Pathogen, Drechslera sorghicola

  • Khan, A.J.;Deadman, M.L.;Al-Maqbali, Y.M.;Al-Sabahi, J.;Srikandakumar, A.;Rizvi, S.G.
    • The Plant Pathology Journal
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    • v.17 no.6
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    • pp.342-346
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    • 2001
  • The physiological changes in sorghum (Sorghum vulgare Pers.) leaves infected with Drechslera sorghicola were investigated through five recognizable stages of disease development. Water-soaked yellowish brown spots developed two days after inoculation, turned brown with yellow halo, enlarged and coalesced at later stages of disease development. Healthy and infected leaves were analyzed for different biochemical constituents. The chlorophyll contents were decreased significantly with the progress of infection. The levels of reducing and total sugars increased while non-reducing sugars decreased to a significant extent with the progress of disease. The concentration of total phenolics, orthodihydroxy phenols, free and glycosidic phenols showed significant changes due to infection, whereas basic and acid phenols showed little or no change with disease development. Levels of phenolic compounds increased four days after inoculation and decrease thereafter, but the concentration was higher at every stage of disease development relative to healthy tissues. Polyphenol oxidase and peroxidase enzyme activities increased to varying degrees at different stages of infection. Analysis of protein fractions showed a significant increase with the progress of disease.

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Molecular Detection and Analysis of Sweet potato feathery motile vims from Root and Leaf Tissues of Cultivated Sweet Potato Plants

  • Ryu, Ki-Hyun;Park, Sun-Hee
    • The Plant Pathology Journal
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    • v.18 no.1
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    • pp.12-17
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    • 2002
  • For the molecular detection of Sweet potaio feathery mottle virus (SPFMV) from diseased sweet potato plants, reverse transcription and polymerase chain reaction (RT-PCR) was performed with the use of a set of virus-specific primers to amplify an 816 bp product. The viral coat protein gene was selected for the design of the primers. No PCR product was amplified when Turnip mosaic virus, Potato vims Y or Cucumber mosaic virus were used as template in RT-PCR with the SPFMV-specific primers. The lowest concentration of template viral RNA required for detection was 10 fg. The vim was rapidly detected from total nucleic acids of leaves and roots from the virus-infected sweet potato plants as well as from the purified viral RNA by the RT-PCR. Twenty-four sweet potato samples were selected and analyzed by RT-PCR and restriction fragment length polymorphism (RFLP). RFLP analysis of the PCR products showed three restriction patterns, which resulted in some point mutations suggesting the existence of quasi-species for the vims in the infected sweet potato plants.

Molecular Analysis of Geminigirus ORFs on Symptom Development

  • Park, Eulyong;Hyunsik Hwang;Lee, Sukchan
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.38-43
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    • 1999
  • Mutants of the monopartite geminivirus beet curly top virus (BCTV) have been screened for infectivity, systemic movement, replication and symptom development in Arabidopsis thaliana. As known by coding for coat protein, R1 mutant was not infectious and did not move systemically. R2, R3 and L2/L3 mutants produced milder symptoms compared to wild type BCTV but the infectivity was reduced by 40% to 60%. R2 ORF is thought to be involved in the regulation of ssDNA and dsDNA accumulation because only dsDNA was accumulated on R2-infected organs. Disruption of ORF L4 resulted in reduced infections, but the viral DNA was accumulated in infected organs from roots to shoot tips as much as wild type BCTV on Sei-O. In addition, 4 mutants did not produce callus-like tissues on infected organs, suggesting that L4 ORF may play a role in the induction of host cell divisions by virus infection. This result was supported by the patterns of mRNA expression and promoter analysis of the cell cycle marker gene, cycl, on Arabidopsis. cycl mRNA was accumulated on symptomatic organs by wild type BCTV infections but not by L4 mutant. We conclude that the BCTV L4 ORF is essential for symptom developments, specially callus-like formation on infected organs.

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Introduction of Hog Cholera Virus Gene into Potato Plants by Agrobacterium-mediated Transformation and the Analysis of Its Expression

  • Kim, Hyun-Soon;Jeon, Jae-Heung;Kim, Cheol-Jung;Hyouk Joung
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.155-161
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    • 2002
  • The HCV gene was expressed in potato plants under the control of the constitutive CaMV 355 promoter or tuber-specific patatin promoter. Solanum tuberosum plants carrying a plant expression vector harboring the encoding region of HCV gene were generated by Agrobacterium tumefaciens-mediated in vitro transformation methods. The presence of HCV gene in the plant genome was detected by PCR and DNA hybridization experiments. We obtained the 5 lines of transgenic potato with the pMBPHCV construct and 4 lines of transgenic potato with the pATHCV construct. The HCV transgenic stably integrated into the potato genome, as well as their transcription. HCV mRNA was identified in leaf and tuber tissues of transgenic plants by Northern blot analysis. The transgenic potato plants produced the expected transcript, and the corresponding HCV protein accumulated in individual transgenic plants.

Expression of IgG1 Germline Transcripts in Germinal Center B Cells of Mouse Popliteal Lymph Nodes

  • Lee, In-Woo;Kim, Jin-Ho;Chung, Gook-Hyun
    • BMB Reports
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    • v.29 no.2
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    • pp.127-132
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    • 1996
  • Germinal centers (GCs) are formed in peripheral lymphoid tissues in response to protein antigens. In order to see if immunoglobulin isotype switching takes place in GC B-cells, we isolated GC B-cells (PNA positive cells) from mouse popliteal lymph nodes by a flow cytometer after the staining of lymph node cells with PNA-FITC and anti-B220-PE, and determined the expression of ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA by RT-PCR. ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA were amplified specifically in cDNAs from hybridoma expressing IgG1 or splenocytes stimulated LPS plus IL-4. Germinal center B-cells formed in popliteal lymph nodes of mice immunized with chicken ovalbumin were isolated 7 days after immunization. We sorted GC B-cells five times. Immunoglobulin ${\gamma}1$ germline transcripts were expressed in germinal center B-cells in three out of five sorts whereas two out of five sorts did not express ${\gamma}1$ germline transcripts in GC B-cells. The contents of GC B-cells ranged from 5 to 7% of total lymph node cells in most flow cytometric analyses but those of two sorted cells which did not express ${\gamma}1$ germline transcripts were out of normal range. These results imply that isotype switching of immunoglobulins may take place in GCs.

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Immobilization and Characterization of a Liposome-Mediated Reconstituted Nicotinic Acetylcholine Receptor

  • Suh, Jeong-Ihn;Palk, Bo-Hyun;Oh, Se-Zu;Suh, Jung-Hun;Cho, Key-Seung;Palk, Young-Ki
    • BMB Reports
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    • v.28 no.2
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    • pp.155-161
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    • 1995
  • A nicotinic acetylcholine receptor (nAchR) isolated from the electric tissues of Torpedo californica has been reconstituted into a vesicle comprising a bifunctional azo-ligand (Bae 1) compound, and a liposome containing phospholipids and cholesterol (1 : 1, w/w). The liposome-mediated reconstituted receptor showed a concentration-dependent response to cholinergic drugs in a lithium ion flux assay. This liposome-mediated reconstituted nAchR was immobilized onto an electrode using various synthetic polymers which were tested for their response to the cholinergic ligands. The immobilized nAchR not only exhibited a linear response to a wide range of cholinergic ligand concentrations but also retained an operational stability which lasted for longer than 6 days. Thus, this result provides a basis for application of the immobilized nAchR-based biosensor in detecting cholinergic ligands in vitro.

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From proteomics toward systems biology: integration of different types of proteomics data into network models

  • Rho, Sang-Chul;You, Sung-Yong;Kim, Yong-Soo;Hwang, Dae-Hee
    • BMB Reports
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    • v.41 no.3
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    • pp.184-193
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    • 2008
  • Living organisms are comprised of various systems at different levels, i.e., organs, tissues, and cells. Each system carries out its diverse functions in response to environmental and genetic perturbations, by utilizing biological networks, in which nodal components, such as, DNA, mRNAs, proteins, and metabolites, closely interact with each other. Systems biology investigates such systems by producing comprehensive global data that represent different levels of biological information, i.e., at the DNA, mRNA, protein, or metabolite levels, and by integrating this data into network models that generate coherent hypotheses for given biological situations. This review presents a systems biology framework, called the 'Integrative Proteomics Data Analysis Pipeline' (IPDAP), which generates mechanistic hypotheses from network models reconstructed by integrating diverse types of proteomic data generated by mass spectrometry-based proteomic analyses. The devised framework includes a serial set of computational and network analysis tools. Here, we demonstrate its functionalities by applying these tools to several conceptual examples.