• Title/Summary/Keyword: endogenous regulator

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Polyamine Stimulation of arcA Expression in Escherichia coli

  • Rhee, Mun-Su;Kim, Young-Sik;Park, Seon-Young;Park, Myung-Hun;Kim, Bo-Min;Kang, Seong-Uk;Lee, Kui-Joo;Lee, Jong-Ho
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.305-312
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    • 2002
  • The effects of two natural polyamines (putrescine and spermidine) on the synthesis of ArcA, a response regulator of the Arc two-component signal transduction system, were studied using an E. coli mutant deficient in polyamine biosynthesis. Endogenous polyamine deficiency of the mutant resulted in marked reduction in the ArcA level determined by Western blot analysis. Putrescine supplement to the growth medium effectively increased the ArcA level of the mutant in a concentration-dependent manner. Spermidine also stimulated the ArcA level in the mutant to a greater degree than putrescine. Expression of arcA'::lacZ operon fusion in the mutant was stimulated 6-fold and 10-fold by putrescine and spermidine at a 1mM concentration, respectively, indicating that the stimulatory effect of the polyamines on ArcA synthesis is due to transcriptional induction, and that spermidine is a more potent arcA inducer than putrescine. The polyamine-dependent arcA'::lacZ induction was growth-phase-dependent and independent of either arcA or fnr which are two regulators involved in anaerobic stimulation of the Arch level. These results suggested that putrescine and spermidine polyamines may be potential intracellular signal molecules in the control of arcA expression, and thereby may play an important role in cellular metabolism.

In vitro shoot initiation of Artocarpus heterophyllus Lam. (Jak Fruit) Effect of the explant type and the season of explant collection

  • Kahk, Kasturiarachchi;Wtpsk, Senarath;Lee, Kui-Jae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10b
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    • pp.2-3
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    • 2003
  • A method for rapid propagation of mature Jack fruit was developed. Four types of explants (mature embryos, apical meristems of young seedlings, apices from mature plants and nodal segments) were used. It has been found 88% of young apical meristems produced shoots in Campbell and Durzan (CD) medium compared to 60% in Murashige and Skoog (MS) medium. Only 1/3 of them produced multiple shoots. Shoot initiation from nodal segments was very rare. Mature apices produced callus. Although removal of the sheathing cover around mature buds enhanced the shoot initiation but success rate was low in growth regulator free medium. Embryos respond to the CD medium but not to the MS medium. Embryos from seeds soaked in water for 24 hours produced shoots after 8 weeks of incubation and the success rate was 70% while embryos from dry seeds only produced roots. There was no significant effect of cold storage (refrigeration) for 7 days on shoot initiation from mature embryos (65%) but the ability for shoot induction declines with storage time (55% after 21 days of cold storage). Mature axillary buds were established in Modified Campbell and Durzan (CD) medium supplemented with 0.5mg/1 and IBA. There was a significant difference in the growth performance of shoots according to the period of the year in which explants were collected. Highest (60%) was observed in November-January period. It was only 30% when the explants were collected in February-April or May-July and decreased to 20% in August-October. The shoots produced in November-January showed a higher vigor than those produced in other months. Since Jak fruit show seasonal changes in fruit bearing and shedding of leaves, it can be suggested that the difference in growth performances of tissues cultured in artificial culture media would have been affected by endogenous rhythms.

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Wnt/$\beta$-catenin/Tcf Signaling Induces the Transcription of a Tumor Suppressor Axin2, a Negative Regulator of the Signaling Pathway

  • Jho, Eek-hoon;Tong Zhang;Claire Domon;Joo, Choun-Ki;Freund, Jean-Noel;Frank Costantini
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.108-108
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    • 2001
  • Axin2/Conductin/Axil and its ortholog Axin are negative regulators of the Wnt signaling pathway, which promote the phosphorylation and degradation of ${\beta}$-catenin. While Axin is expressed ubiquitously, Axin2 mRNA was seen in a restricted pattern during mouse embryogenesis and organogenesis. Because many sites of Axin2 expression overlapped with those of several Wnt genes, we tested whether Axin2 was induced by Wnt signaling. Endogenous Axin2 mRNA and protein expression could be rapidly induced by activation of the Wnt pathway, and Axin2 reporter constructs, containing a 5.6 kb DNA fragment including the promoter and first intron, were also induced. This genomic region contains eight Tcf/LEF consensus binding sites, five of which are located within longer, highly conserved non-coding sequences. The mutation or deletion of these Tcf/LEF sites greatly diminished induction by ${\beta}$-catenin, and mutation of the Tcf/LEF site T2 abolished protein binding in an electrophoretic mobility-shift assay. These results strongly suggest that Axin2 is a direct target of the Wnt pathway, mediated through Tcf/LEF factors. The 5.6 kb genomic sequence was sufficient to direct the tissue specific expression of d2EGFP in transgenic embryos, consistent with a role for the Tcf/LEF sites and surrounding conserved sequences in the in vivo expression pattern of Axin2. Our results suggest that Axin2 participates in a negative feedback loop, which could serve to limit the duration or intensity of a Wnt-initiated signal.

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NELL2 Function in Axon Development of Hippocampal Neurons

  • Kim, Han Rae;Kim, Dong Hee;An, Ji Young;Kang, Dasol;Park, Jeong Woo;Hwang, Eun Mi;Seo, Eun Jin;Jang, Il Ho;Ha, Chang Man;Lee, Byung Ju
    • Molecules and Cells
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    • v.43 no.6
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    • pp.581-589
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    • 2020
  • Neurons have multiple dendrites and single axon. This neuronal polarity is gradually established during early processes of neuronal differentiation: generation of multiple neurites (stages 1-2); differentiation (stage 3) and maturation (stages 4-5) of an axon and dendrites. In this study, we demonstrated that the neuron-specific n-glycosylated protein NELL2 is important for neuronal polarization and axon growth using cultured rat embryonic hippocampal neurons. Endogenous NELL2 expression was gradually increased in parallel with the progression of developmental stages of hippocampal neurons, and overexpression of NELL2 stimulated neuronal polarization and axon growth. In line with these results, knockdown of NELL2 expression resulted in deterioration of neuronal development, including inhibition of neuronal development progression, decreased axon growth and increased axon branching. Inhibitor against extracellular signal-regulated kinase (ERK) dramatically inhibited NELL2-induced progression of neuronal development and axon growth. These results suggest that NELL2 is an important regulator for the morphological development for neuronal polarization and axon growth.

Effect of Polyamine on Hairy Root Culture of Bupleurum falcatum L. (Polyamine이 시호의 모상근 배양에 미치는 영향)

  • 표병식;김영준황백
    • KSBB Journal
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    • v.10 no.4
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    • pp.435-440
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    • 1995
  • During culture the effect of polyamine on hairy root of Buplerum falcatum by infection of Agrobacterium rhizogenes was studied. The fresh and dry weight of hairy root which cultured for 3months in MS medium increased about 7-fold when spermine at 10${\mu}$M, 100${\mu}$M was treated. After suspension culture of B. falcatum in MS medium containing putrescine(10${\mu}$M, 100${\mu}$M) or spermine(10${\mu}$M), the contents of endogenous polyamine (putresclne, spermidine, spermine) was higher than that of control. The ${\beta}$-glucan synthetase II activity by polyamine treatment was increased: especially spermidine (100${\mu}$M) and spermine(100${\mu}$M) stimulated it by about 180% and 220% respectively. These results suggest a possible role of polyamine as growth regulator in B. falcatum hairy root cultures.

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Evidence for Existence of a Water-Extractable Anticoagulant in an Earthworm, Lumbricus rubellus

  • Woo, Jeong-Im;Bahk, Yun-Kyung;Yu, Kyoung-Hee;Paik, Seung-R.;Chang, Chung-Soon
    • BMB Reports
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    • v.29 no.6
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    • pp.500-506
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    • 1996
  • We have isolated a water-extracted novel regulator for blood coagulation from an earthworm, Lumbricus rubellus. As a folk remedy, the earthworm has been known to facilitate blood circulation. After complete heat inactivation of endogenous proteases in the earthworm, an anticoagulant(s) was purified through ammonium sulfate fractionation and three consecutive gel permeation chromatography of Sephacryl S-300, Sephadex G-75, and G-150 by measuring activated partial thromboplastin time (APTT) The anticoagulant was further purified to 2,800 fold with a C4 reversed-phase HPLC This activity was stable under heat ($100^{\circ}C$ for 30 min) and acidic conditions (0.4 N HCl). The effects of this partially purified anticoagulant on thrombin were observed with various substrates such as N${\alpha}$-benzoyl-DL-arginine-p-nitroanilide (BApNA), H-D-phenylalanyl-L-pipecoyl-L-arginine-p-nitroanilide (S-2238), N${\alpha}$-p-tosyl-L-arginine methyl ester (TAME), and fibrinogen as a natural substrate. Only TAME hydrolysis, due to an esterase activity of the enzyme, was inhibited among the chromogenic substrates. In addition, the anticoagulant not only inhibited the conversion of fibrinogen to fibrin but also prolonged the fibrin clot formation monitored with the in vitro coagulation test. Based on these observations, we suggest the significance of measuring the ability of antithrombotic drugs to inhibit the esterase activity of thrombin. In this report, it was also shown that the earthworm indeed contained a water-extractable, heat- and acid-stable anticoagulant which could be used as a novel antithrombotic agent.

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In vitro shoot initiation of Artocarpus heterophyllus Lam. (Jak Fruit) Effect of the explant type and the season of explant collection

  • Kahk, Kasturiarachchi;Wtpsk, Senarath;Lee, Kui-Jae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10a
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    • pp.9-18
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    • 2003
  • A method for rapid propagation of mature Jack fruit was developed. Four types of explants (mature embryos, apical meristems of young seedlings, apices from mature plants and nodal segments) were used. It has been found 88% of young apical meristems produced shoots in Campbell and Durzan (CD) medium compared to 60% in Murashige and Skoog (MS) medium. Only 1/3 of them produced multiple shoots. Shoot idtiation from nodal segments was very rare. Mature apices produced callus. Although removed of the sheathing cover around mature buds enhanced the shoot initiation but success rate was low in growth regulator free medium. Embryos respond to the CD medium but not to the MS medium. Embryos from seeds soaked in water for 24 hours produced shoots after 8 weeks of incubation and the success rate was 70% while embryos from dry seeds only produced roots. There was no significant effect of cold storage (refrigeration) for 7 days on shoot initiation from mature embryos (65%) but the ability for shoot induction declines with storage time (55% after 21 days of cold storage). Mature axillary buds were established in Modified Campbell and Durzan (CD) medium supplemented with 0.5mg/1 and IBA. There was a significant difference in the growth performance of shoots according to the period of the year in which explants were collected. Highest (60%) was observed in November-January period. It was only 30% when the explants were collected in February-April or May-July and decreased to 20% in August-October. The shoots produced in November-January showed a higher vigor than those produced in other months. Since Jak fruit show seasonal changes in fruit bearing and shedding of leaves, it can be suggested that the difference in growth performances of tissues cultured in artificial culture media would have been affected by endogenous rhythms.

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Similarities and Distinctions in the Effects of Metformin and Carbon Monoxide in Immunometabolism

  • Park, Jeongmin;Joe, Yeonsoo;Ryter, Stefan W.;Surh, Young-Joon;Chung, Hun Taeg
    • Molecules and Cells
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    • v.42 no.4
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    • pp.292-300
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    • 2019
  • Immunometabolism, defined as the interaction of metabolic pathways with the immune system, influences the pathogenesis of metabolic diseases. Metformin and carbon monoxide (CO) are two pharmacological agents known to ameliorate metabolic disorders. There are notable similarities and differences in the reported effects of metformin and CO on immunometabolism. Metformin, an anti-diabetes drug, has positive effects on metabolism and can exert anti-inflammatory and anti-cancer effects via adenosine monophosphate-activated protein kinase (AMPK)-dependent and AMPK-independent mechanisms. CO, an endogenous product of heme oxygenase-1 (HO-1), can exert anti-inflammatory and antioxidant effects at low concentration. CO can confer cytoprotection in metabolic disorders and cancer via selective activation of the protein kinase R-like endoplasmic reticulum (ER) kinase (PERK) pathway. Both metformin and CO can induce mitochondrial stress to produce a mild elevation of mitochondrial ROS (mtROS) by distinct mechanisms. Metformin inhibits complex I of the mitochondrial electron transport chain (ETC), while CO inhibits ETC complex IV. Both metformin and CO can differentially induce several protein factors, including fibroblast growth factor 21 (FGF21) and sestrin2 (SESN2), which maintain metabolic homeostasis; nuclear factor erythroid 2-related factor 2 (Nrf2), a master regulator of the antioxidant response; and REDD1, which exhibits an anticancer effect. However, metformin and CO regulate these effects via different pathways. Metformin stimulates p53- and AMPK-dependent pathways whereas CO can selectively trigger the PERK-dependent signaling pathway. Although further studies are needed to identify the mechanistic differences between metformin and CO, pharmacological application of these agents may represent useful strategies to ameliorate metabolic diseases associated with altered immunometabolism.

RAV1 Negatively Regulates Seed Development by Directly Repressing MINI3 and IKU2 in Arabidopsis

  • Shin, Hyun-young;Nam, Kyoung Hee
    • Molecules and Cells
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    • v.41 no.12
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    • pp.1072-1080
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    • 2018
  • A plant-specific B3 domain and AP2 domain-containing transcription factor, RAV1 acts as a negative regulator of growth in many plant species and its transcription was down-regulated by BR and ABA. In this study, we found that RAV1-overexpressing transgenic plants showed abnormally developed ovules, resulting in reduced seed size, weight, and number in a silique. Interestingly, the endogenous expression of RAV1 fluctuated during seed development; it remained low during the early stage of seed development and sharply increased in the seed maturation stage. In plants, seed development is a complex process that requires coordinated growth of the embryo, endosperm, and maternal integuments. Among many genes that are associated with endosperm proliferation and embryo development, three genes consisting of SHB1, MINI3, and IKU2 form a small unit positively regulating this process, and their expression was regulated by BR and ABA. Using the floral stage-specific RNAs, we found that the expression of MINI3 and IKU2, the two downstream genes of the SHB1-MINI3-IKU2 cascade in the seed development pathway, were particularly reduced in the RAV1-overexpressing transgenic plants. We further determined that RAV1 directly binds to the promoter of MINI3 and IKU2, resulting in their repression. Direct treatment with brassinolide (BL) improved seed development of RAV1-overexpressing plants, but treatment with ABA severely worsened it. Overall, these results suggest that RAV1 is an additional negative player in the early stages of seed development, during which ABA and BR signaling are coordinated.

Interaction between Brassinolide and Auxins on Bioassays (Brassinolide와 Auxin과의 상호작용의 생물검정에 의한 평가)

  • Choi, Chung-Don;Kim, Soon-Chul;Lee, Soo-Kwan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.1
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    • pp.58-64
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    • 1990
  • This study was conducted to investigate the interaction effect of a new plant growth regulator brassinolide and auxins using several bioassay methods such as rice lamina inclination test, oat coleoptile segment growth test and radish hypocotyl elongation test. For rice lamina inclination test, the antagonistic response showed at low concentration combinations of homobrassinolide and auxins (IAA and 2,4-D), while the combinations of high concentration responsed mostly synergistic or additive effects, IAA was generally higher than 2,4-D for combination effect with homobrassinolide. For oat coleoptile segment growth test, the synergistic effect showed at IAA concentrations less than 0.1ppm while additive response exhibited above 0.3ppm regardless of homobrassinolide concentrations. In radish hypocotyl elongation test, the interaction response varied with cotyledon. The sections removed cotyledon showed mostly antagonistic effects, except for combination of homobrassinolide with IAA 3ppm. Interaction responses were Quite different from bioassay methods, particularly using experimental materials: antagonistic responses were usually shown at the section that attached growing point while these for sections that removed growing point were responded synergistic or additive effect due to unstable endogenous hormones.

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