• Title/Summary/Keyword: metallothionein expression

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OsWRKY42 Represses OsMT1d and Induces Reactive Oxygen Species and Leaf Senescence in Rice

  • Han, Muho;Kim, Chi-Yeol;Lee, Junok;Lee, Sang-Kyu;Jeon, Jong-Seong
    • Molecules and Cells
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    • v.37 no.7
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    • pp.532-539
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    • 2014
  • We isolated a rice (Oryza sativa L.) WRKY gene which is highly upregulated in senescent leaves, denoted OsWRKY42. Analysis of OsWRKY42-GFP expression and its effects on transcriptional activation in maize protoplasts suggested that the OsWRKY42 protein functions as a nuclear transcriptional repressor. OsWRKY42-overexpressing (OsWR KY42OX) transgenic rice plants exhibited an early leaf senescence phenotype with accumulation of the reactive oxygen species (ROS) hydrogen peroxide and a reduced chlorophyll content. Expression analysis of ROS producing and scavenging genes revealed that the metallothionein genes clustered on chromosome 12, especially OsMT1d, were strongly repressed in OsWRKY42OX plants. An OsMT1d promoter:LUC construct was found to be repressed by OsWRKY42 overexpression in rice protoplasts. Finally, chromatin immunoprecipitation analysis demonstrated that OsWRKY42 binds to the W-box of the OsMT1d promoter. Our results thus suggest that OsWRKY42 represses OsMT1d-mediated ROS scavenging and thereby promotes leaf senescence in rice.

Differential Gene Expression in Estradiol-3-Benzoate-Treated Liver and Chemically- Induced Hepatocellular Carcinoma

  • KIM , SEYL;KANG, JIN-SEOK;JANG, DONG-DEUK;LEE, KOOK-KYUNG;KIM, SOON-AE;HAN, BEOM-SEOK;PARK, YOUNG-IN
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1286-1294
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    • 2004
  • In a previous study by the current authors, hepatocellular carcinoma (HCC) was determined to be epidemiologically sex-dependent, and the incidence and multiplicity of HCC found to decrease in estradiol-3 benzoate (EB)-treated F344 rats. Therefore, to ascertain the anticancer mechanism of EB, a commercially available cDNA microarray, with a total of 14,815 cDNA rat gene clones, was used to determine the differentially expressed genes in nontreated livers, EB-treated livers, and diethynitrosolamine (DEN)-induced HCC. In the sequenced experiment, a total of 85 genes were differentially expressed at either two or more times the rate of the normal expression, where 33 genes were downregulated by EB, and 52 genes upregulated. Candidate genes were selected according to significant changes observed in the mRNA expression in the EB-treated livers compared with the nontreated livers, then these genes were filtered according to their different expression patterns in the DEN-induced tumors compared to the estrogen-treated livers. To confirm the microarray data, a real-time PCR analysis was performed for ten selected genes: the H-ras revertant protein 107 (H­rev107), insulin-like growth factor binding protein (lOFBP), parathyroid hormone receptor (PI'HR), SH3 domain binding protein (SH3BP), metallothionein, src-suppressed C-kinase substrate (SSeCK) gene, phosphodiesterase I, CD44, epithelial membrane protein 3 (EMP3), and estrogen receptor a (ERa). The SSeCK and phosphodiesterase I genes were both upregulated in the DEN-induced hepatocarcinomas, yet their possible carcinogenic functions remain unknown. Meanwhile, the other genes were downregulated, including the genes related to growth regulation (IOFBP, H-revI07, ER$\alpha$), adipogenesis inhibition (PTHR), and tumor suppression (metallothionein).

Heavy-Metal Adsorption by Recombinant Saccharomyces cerevisiae Harboring Multiple Copies of the CUP1 Gene (구리흡착 단백질 유전자를 함유하는 재조합 효모의 중금속 흡착)

  • 서진호;박상옥;김명동;한기철;전영석;안장우;한남수
    • KSBB Journal
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    • v.17 no.1
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    • pp.38-43
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    • 2002
  • Characteristics of cell growth and heavymetal adsorption by recombinant Saccharomyus cerevisiae strains harboring multiple copies of the CUP1 gene encoding metallothione (MT) protein were studied in batch cultures. Recombinant S. cerevisiae strains harboring multiple copies of the CUP1 gene were superior to the host and wild-type yeast strains in terms of cell growth and heavy metal removal, indicating that the copy number of the CUP1 gene for MT expression played an important role in the adsorption of heavy metals. It was suggested that the CUP1 promoter for the MT expression is induced by manganese and zinc as well as copper An optimum copper concentration for MT expression and concomitant adsorption of heavy metals by recombinant S. cerevisiae was found to be 0.31 mM. A nonionic surfactant Triton X-100 enhanced cell growth by 17.7% and removal of zinc by 6.1% compared with the control case.

Characteristics of Growth and Metal Removal in Recombinant Saccharomyces cerevisiae harboring a Metallothionein Gene (Metallothionein 유전자가 도입된 재조합 Saccharomyces cerevisiae의 생육과 금속제거에 대한 특성)

  • 정동환;김대옥서진호
    • KSBB Journal
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    • v.10 no.5
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    • pp.475-481
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    • 1995
  • The effect of metallothionein expression on the metal resistance and removal by recombinant Saccharomyces cerevisiae containing the plasmid pJW9 was investigated. The recombinant strain S. cerevisiae BZ-pJ was constructed by transforming the host strain S. cerevisiae BZ3l-1-7Ba with the gene coding for a metal-binding protein, metallothionein. Introduction of the MT gene yielded an increase in the minimum inhibitory concentration (MIC) of copper more than three times compared with the host strain. The minimum inhibitory concentrations of $Cr^{2+}, Znr^{2+} and Pb^{2+}, $ were not different for the two strains. The recombinant yeast grown in a medium containing 8mM CuSO4 was able to remove copper with a capacity of 18.9mg $Cu^{2+}$/g dry cell. In a mixture of copper and zinc, the presence of copper relieved the toxic effects caused by zinc, resulting in an enhancement of the final cell density and the specific growth rate of the recombinant yeast. The capability to remove copper by the recombinant yeast was linearly proportional to the copper concentrations in the medium. The efficiency of copper removal was rather constant regardless of the initial copper concentrations. The specific removal of zinc was dependent on the zinc concentrations in media, though, and such dependence was not so pronounced as the concentration of copper.

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The Production of Foreign Protein in Mouse L Cell (Mouse L Cell에서의 외래 유전자 유래 단백질의 생산)

  • ;Tadamitsu Kishimoto
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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    • pp.421-427
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    • 1993
  • Some interleukin 6 (IL-60 transcription control factors were resported as the regulator of IL-6 expression. A nuclear protein bound to interleukin 1 (IL-1) responsive element in the IL-6 promoter region was named NF-IL6 (nuclear factor for IL-6). This NF-IL6 was known to be very imporant as a transcription factor for various immuno-protein as well as for IL-6. The human NF-IL6 genes were transfected into the mouse L cells under the metallothionein promoter (MT promoter) to establish a model system for the expression of foreign gene in the mammalian cell line.

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Zinc Status Assessment by Analysis of Mononuclear Cell Metallothionein mRNA Using Competitive-Reverse Transcriptase-Polymerase Chain Reaction

  • Lee, Soo-Lim;Yoon, Jin-Sook;Kwon, Chong-Suk;Beattie, John H.;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.9 no.3
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    • pp.276-282
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    • 2004
  • Marginal Zn deficiency is prevalent through the world and yet human zinc status has not been properly assessed due to the lack of a reliable diagnostic indicator. One potential possibility for zinc status assessment using Zn-binding protein, metallothionein (MT)-mRNA, has been proposed. The purpose of the present study was aimed to show whether measurement of mononuclear cell (MNC) MT mRNA, using a competitive-reverse transcriptase-polymerase chain reaction (competitive-RT-PCR) assay, could indicate zinc status in human subjects. In this study, MNC MT-mRNA expression was measured using a competitive-RT-PCR to compare before and after 14 days of zinc supplementation (50 mg Zn/das zinc gluconate). RT-PCR oligonucleotide primers which were designed to amplify both a 278 bp segment of the human MT-2A cDNA and a 198 bp mutant competitor cDNA template from MNCs, were prepared. MT-2A mRNA was normalized by reference to the housekeeping gene, $\beta$-actin, mRNA for which was also measured by competitive-RT-PCR. There was considerable inter-individual variation in MT-mRNA concentration and yet, the mean MT-2A mRNA level increased 4.7-fold after Zn supplementation, as compared to before Zn supplementation. This MT-2A mRNA level was shown as the same pattern and, even more sensitive assay, compared to the conventional plasma and red blood cells (RBCs) Zn assessment in which plasma and RBCs zinc levels increased 2.3- and 1.2-fold, respectively (p<0.05). We suggest that MT competitive-RT-PCR can be a useful assessment tool for evaluating human zinc status.