• Title/Summary/Keyword: soxB

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Regulation of SoxR, the superoxide-sensory regulator in Escherichia coli.

  • Lee Joon-Hee;Koo Mi-Sun;Yeo Won-Sik;Roe Jung-Hye
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.24-31
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    • 2000
  • In order to find out SoxR-reducing system in E. coli, we generated Tn10-insertion mutants and screened for constitutive expression of SoxS in a soxS-lacZ fusion strain. One mutation was mapped in rseB, a gene in rseABC (Regulation of SigmaE) operon. The constitutive soxS-expressing phenotype was due to the polar effect on the downstream gene, rseC. RseC is likely to function as a component of SoxR reduction system because SoxR was kept in oxidized form to activate soxS expression in rseC mutant. RseC is an integral membrane protein with an N-terminal cysteine-rich domain in the cytoplasm. The functionally critical cysteines were determined by substitution mutagenesis. The truncated N-terminal domain of RseC reduced the soxS transcription by $50\%$ as judged by in vitro transcription assay. Currently RseC is believed to be a reducing factor for SoxR. However, the mechanism for the reduction needs further investigation.

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DDX53 Regulates Cancer Stem Cell-Like Properties by Binding to SOX-2

  • Kim, Youngmi;Yeon, Minjeong;Jeoung, Dooil
    • Molecules and Cells
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    • v.40 no.5
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    • pp.322-330
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    • 2017
  • This study investigated the role of cancer/testis antigen DDX53 in regulating cancer stem cell-like properties. DDX53 shows co-expression with CD133, a marker for cancer stem cells. DDX53 directly regulates the SOX-2 expression in anti-cancer drug-resistant $Malme3M^R$ cells. DDX53 and miR-200b were found to be involved in the regulation of tumor spheroid forming potential of Malme3M and $Malme3M^R$ cells. Furthermore, the self-renewal activity and the tumorigenic potential of $Malme3M^R$-CD133 (+) cells were also regulated by DDX53. A miR-200b inhibitor induced the direct regulation of SOX-2 by DDX53 We therefore, conclude that DDX53 may serve as an immunotherapeutic target for regulating cancer stem-like properties of melanomas.

무당개구리 (Bombina orientalis) 배아에서 제초제 Molinate에 의한 발생 위해성 평가 및 Biomarker 유전자 발굴

  • Gang Han Seung;Gye Myeong Chan;Kim Mun Gyu
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2004.12a
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    • pp.124-126
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    • 2004
  • Molinate is thiocarbamate herbicide used primarily in rice production. Chondrogenesis is a multistep process that is essential for endocondral bone formation. The transcription factor Sox9 has an essential role during the sequential steps of chondrocyte differentiation. Bombina orientalis is one of the most common amphibians in the world and comprises a large proportion of their total number. We examined the embryotoxic and survival effects of molinate at various concentration in B. orientalis embryos. The survival rates of embryos at 312h post fertilization treated with molinate was decreased with concentration dependent manner. Also, developmental malformations appeared by molinate in B. orientalis embryos. the expression levels of Sox9 mRNA was examined by RT-PCR. In our result showed that Sox9 expression was found to be increased in malformed tadpole compared to normal tadpole. These results suggested that molinate was detrimental for survival and development of B. orientalis embryo.

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A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

The Virulence of Vibrio vulnificus is Affected by the Cellular Level of Superoxide Dismutase Activity

  • Kang, In-Hye;Kim, Ju-Sim;Lee, Jeong-K.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1399-1402
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    • 2007
  • The virulence of superoxide dismutase (SOD) mutants of Vibrio vulnificus, as tested by intraperitoneal injection into mice, decreases in the order of sodC mutant, sodA mutant, and sodB mutant lacking CuZnSOD, MnSOD, and FeSOD, respectively. The survival of SOD mutants under superoxide stress also decreases in the same order. The virulence of soxR mutant, which is unable to induce MnSOD in response to superoxide, is similar to that of the sodA mutant, as the survival of the soxR mutant under superoxide stress is similar to that of the sodA mutant. Consistently, the lowered survival of the soxR mutant is complemented not only with soxR but also with sodA. Thus, the virulence of V. vulnificus is significantly affected by the cellular level of SOD activity, and an increase in SOD level through MnSOD induction by SoxR under superoxide stress is essential for virulence.

Isolation and Characterization of Paraquat-inducible Promoters from Escherichia coli

  • Lee, Joon-Hee;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.277-283
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    • 1997
  • Promoters inducible by paraquat, a superocide-generating agent, were isolated from Escherichia coli using a promoter-probing plasmid pRS415 with promoterless lacA gene. Twenty one promoters induced by paraquat were selected and further characterized. From sequence analysis, thirteen of the promoters were mapped to their specific loci on the Escherichia coli chromosome. Several promoters were mapped to the upstream of known genes such as usgl, katG, and mglB, whose relationships with superoxide response have not been previously reported. Other promoters were mapped to the upstream region of unknown open reading frames. Downstream of HC 96 promoter are uncharacterized ORFs whose sequences are homologous to ABC-transporter subunits. Downstream of HC84 promoter is an ORF encoding a transcriptional regulator-like protein, which contains a LysR family-specific HTH (helix-turn-helix) DNA bindign motif. We investigated whether these promoters belong to the soxRS regulon. All promoters except HC96 were found to belong to the soxRS regulon. The HC96 promoter was significantly induced by paraquat in the soxRS deletion mutant strain. The basal transcription level of three promoters (HE43, HC71, HD94) significantly increased at the stationary phase, implying that they are regulated by RpoS. However, paraquat inducibility of all promoters disappeared in the stationary phase, suggesting that SoxRS regulatory system is active only in rapidly growing cells.

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Complete genome sequence of Comamonas sp. NLF-7-7 isolated from biofilter of wastewater treatment plant (폐수처리장의 바이오 필터로부터 분리된 Comamonas sp. NLF-7-7 균주의 유전체 염기서열 해독)

  • Kim, Dong-Hyun;Han, Kook-Il;Kwon, Hae Jun;Kim, Mi Gyeong;Kim, Young Guk;Choi, Doo Ho;Lee, Keun Chul;Suh, Min Kuk;Kim, Han Sol;Lee, Jung-Sook;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.309-312
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    • 2019
  • Comamonas sp. NLF-7-7 was isolated from biofilter of wastewater treatment plant. The whole-genome sequence of Comamonas sp. NLF-7-7 was analyzed using the PacBio RS II and Illumina HiSeqXten platform. The genome comprises a 3,333,437 bp chromosome with a G + C content of 68.04%, 3,197 total genes, 9 rRNA genes, and 49 tRNA genes. This genome contained pollutants degradation and floc forming genes such as sulfur oxidization pathway (SoxY, SoxZ, SoxA, and SoxB) and floc forming pathway (EpsG, EpsE, EpsF, EpsG, EpsL, and glycosyltransferase), respectively. The Comamonas sp. NLF-7-7 can be used to the purification of wastewater.

Subpopulations of miniature pig mesenchymal stromal cells with different differentiation potentials differ in the expression of octamer-binding transcription factor 4 and sex determining region Y-box 2

  • Jeon, Ryounghoon;Park, Sungjo;Lee, Sung-Lim;Rho, Gyu-Jin
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.515-524
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    • 2020
  • Objective: Human mesenchymal stromal cells (MSCs) exhibit variable differentiation potential and can be divided accordingly into distinct subpopulations whose ratios vary with donor age. However, it is unknown whether the same is true in pigs. This study investigated MSC subpopulations in miniature pig and compared their characteristics in young (2 to 3 months) and adult (27 to 35 months) pigs. Methods: Osteogenic, chondrogenic, and adipogenic capacity of isolated MSCs was evaluated by von Kossa, Alcian blue, and oil red O staining, respectively. Cell surface antigen expression was determined by flow cytometry. Proliferative capacity was assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Expression of marker genes was detected by quantitative real-time polymerase chain reaction. Results: Porcine MSCs comprised cells with trilineage and bilineage differentiation potential (tMSCs and bMSCs, respectively) and non-differentiating stromal cells (NDSCs). The tMSC and bMSC fractions were smaller in adult than in young pigs (63.0% vs 71.2% and 11.6% vs 24.0%, respectively, p<0.05); NDSCs showed the opposite trend (25.4% vs 4.8%; p<0.05). Subpopulations showed no differences in morphology, cell surface antigen expression, or proliferative capacity, but octamer-binding transcription factor 4 (OCT4) expression was higher in tMSCs than in bMSCs and NDSCs (p<0.05), whereas sex determining region Y-box 2 (SOX2) expression was higher in tMSCs and bMSCs than in NDSCs (p<0.05). Aging had no effect on these trends. Conclusion: Porcine MSCs comprise distinct subpopulations that differ in their differentiation potential and OCT4 and SOX2 expression. Aging does not affect the characteristics of each subpopulation but alters their ratios.

An Evaluation on the Combustion Characteristics of Heavy Oil-Water Emulsions (중질유-물 유화연료의 연소특성 평가)

  • Lee, Yong-Il
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.26 no.12
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    • pp.1722-1728
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    • 2002
  • Most researches regarding emulsified fuel were in the areas of emulsifier, emulsified fuel manufacturing and emulsified fuel droplet combustion, but there were little papers published regarding emulsified fuel combustion and boiler efficiency in an industrial boiler. The main purpose of this study is to clarify whether improvements in the boiler efficiency and the reduction of pollutants such as CO, NOx, SOx and smoke exist or not when emulsified fuels are combusted in the commercial boiler. Main experimental parameters were water content in heavy oil , excess $O_2$, and boiler load. The fuels used in this experiment were 0.5 B-C, and 5 kinds of 0.5 B-C/water emulsified fuels. The combustion characteristics of heavy oil and its emulsions with water were investigated in an industrial boiler. The combustion stability was monitored and exhaust gases such as CO, NOx, SOx and smoke were measured with excess $O_2$ and combustion load. In case of emulsified fuel combustion, flame stability was poor and boiler efficiency was lowered by 1.6~5.7%, but emission levels of CO and smoke were improved.

The Expression Patterns of Cdc25A, Cdc25B, Sox2 and Mnb in Central Nervous System in Early Chicken Embryos

  • Zhang, Hui;Qin, Junhui;Cao, Jingjing;Hei, Nainan;Xu, Chunsheng;Yang, Ping;Liu, Haili;Chu, Xiaohong;Bao, Huijun;Chen, Qiusheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.6
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    • pp.781-787
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    • 2009
  • The sense and antisense digoxigenin-labeled RNA probes of four genes, Cdc25A, Cdc25B, Sox2 and Mnb, were produced by using SP6 and T7 RNA polymerases, respectively, and in vitro transcription. Expression patterns of the four genes were detected by in situ hybridization in HH (Hamburger and Hamilton) stage 10 chick embryos. In general, expression patterns of the four genes were similar. mRNA of the four genes was mostly restricted to the entire CNS (central nervous system). All were confined to an identical region, neural tube, neural groove and caudal neural plate, corresponding to the notochord or spinal cord, but there was some distinction in specific region or in concentration, for example in somites. The overlap in expression at the same developmental stage in the CNS suggests that the four genes may be functional similar or related in CNS development. Expression patterns of the four genes support specific roles of these regulators in the developing CNS.