• Title/Summary/Keyword: ynk mutant

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Two-dimensional Electrophoretic Analysis of Nucleotide phosphate Kinase Mediated Hydrogen Peroxide Cross-linking in Saccharamyces cerevisiae (2-D 전기영동 분석을 통한 $H_2O_2$와 연계된 효모 시스템 NDPK에 관한 연구)

  • Moon Hae-Jeong;Yun Dae-Jin;Park Chang-Ho
    • KSBB Journal
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    • v.21 no.1 s.96
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    • pp.16-19
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    • 2006
  • Oxidative modification of nucleoside diphosphate kinase (NDPK) is identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometer. The quaternary structure of NDPK appears to be regulated by cross-linking with an oxidant, $H_2O_2$. We compared roles of NDPK in each of wild type and ynk mutant against oxidative stress. Six specific proteins changed by $H_2O_2$ were identified using two-dimensional electrophoretic analysis. YNK regulated several proteins, related to $H_2O_2$ signaling functions. These results suggest that one of the important functions of NDPK is the regulation of cellular redox state.

Pro-Apoptotic Role of the Human YPEL5 Gene Identified by Functional Complementation of a Yeast moh1Δ Mutation

  • Lee, Ji Young;Jun, Do Youn;Park, Ju Eun;Kwon, Gi Hyun;Kim, Jong-Sik;Kim, Young Ho
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.633-643
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    • 2017
  • To examine the pro-apoptotic role of the human ortholog (YPEL5) of the Drosophila Yippee protein, the cell viability of Saccharomyces cerevisiae mutant strain with deleted MOH1, the yeast ortholog, was compared with that of the wild-type (WT)-MOH1 strain after exposure to different apoptogenic stimulants, including UV irradiation, methyl methanesulfonate (MMS), camptothecin (CPT), heat shock, and hyperosmotic shock. The $moh1{\Delta}$ mutant exhibited enhanced cell viability compared with the WT-MOH1 strain when treated with lethal UV irradiation, 1.8 mM MMS, $100{\mu}M$ CPT, heat shock at $50^{\circ}C$, or 1.2 M KCl. At the same time, the level of Moh1 protein was commonly up-regulated in the WT-MOH1 strain as was that of Ynk1 protein, which is known as a marker for DNA damage. Although the enhanced UV resistance of the $moh1{\Delta}$ mutant largely disappeared following transformation with the yeast MOH1 gene or one of the human YPEL1-YPEL5 genes, the transformant bearing pYES2-YPEL5 was more sensitive to lethal UV irradiation and its UV sensitivity was similar to that of the WT-MOH1 strain. Under these conditions, the UV irradiation-induced apoptotic events, such as FITC-Annexin V stainability, mitochondrial membrane potential (${\Delta}{\psi}m$) loss, and metacaspase activation, occurred to a much lesser extent in the $moh1{\Delta}$ mutant compared with the WT-MOH1 strain and the mutant strain bearing pYES2-MOH1 or pYES2-YPEL5. These results demonstrate the functional conservation between yeast Moh1 and human YPEL5, and their involvement in mitochondria-dependent apoptosis induced by DNA damage.