• Title/Summary/Keyword: cellular growth

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Growth-Dependent Variations in Antioxidant and Redox Enzyme Activities of Schizosaccharomyces pombe

  • Cho, Young-Wook;Park, Eun-Hee;Ahn, Ki-Sup;Kim, Dae-Myung;Lim, Chang-Jin
    • BMB Reports
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    • v.34 no.3
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    • pp.278-283
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    • 2001
  • Antioxidant and redox enzyme activities are known to be involved in the cellular responses to various stresses. Their variations were observed according to the growth cycle of the fission yeast Schizosaccharomyces pombe. Peroxidase activity appeared to be notably higher in the early exponential phase than in the mid-exponential and stationary phases. However, catalase activity showed a variation pattern resembling the growth curve. Glutathione S-transferase activity was higher in the early exponential and late stationary phases. Activities of the two redox enzymes, thioredoxin and thioltransferase (glutaredoxin), were high in the stationary phase. However, their activities appeared to increase from the early exponential to mid-exponential phase. Total glutathione content had a varying pattern similar to that of thioredoxin and thioltransferase. However, its content in the early exponential phase was high. These results propose that antioxidant and redox enzymes tested are also involved in the mechanism of cell growth.

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Proteases and Protease Inhibitors Produced in Streptomycetes and Their Roles in Morphological Differentiation

  • KIM DAE WI;KANG SUNG GYUN;KIM IN SEOP;LEE BYONG KYU;RHO YONG TAIK;LEE KYE JOON
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.5-14
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    • 2006
  • Streptomycetes are Gram-positive microorganisms producing secondary metabolites through unique physiological differentiation [4]. The microbes show unusual morphological differentiation to form substrate mycelia, aerial mycelia, and arthrospores on solid medium [19]. Substrate mycelium growth is sustaining with sufficient nutrients in the culture medium. The concentration of a specific individual substrate in the culture environment is the most important extracellular factor allowing vegetative mycelia growth, where extracellular hydrolytic enzymes participate in the utilization of waterinsoluble substrates. However, with starvation of nutrients in the culture medium, the vegetative mycelia differentiate to aerial mycelia and spores. It has been considered that shiftdown of essential nutrients for mycelia growth is the most important factor triggering morphological and physiological differentiation in Streptomyces spp. Since proteineous macromolecule compounds are the major cellular components, these are faced to endogenously metabolize following a severe depletion of nitrogen source in culture nutrients (Fig. 1). Various proteases were identified of which production was specifically related with the phase of mycelium growth and also morphological differentiation. The involvement of proteases and protease inhibitor is reviewed as a factor explaining the mycelium differentiation in Streptomyces spp.

Identification of the Arabidopsis thaliana cell growth defect factor suppressing yeast cell proliferation

  • Kim, Kyung-Min;Uchimiya, Hirofumi;Sohn, Jae-Keun
    • Current Research on Agriculture and Life Sciences
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    • v.30 no.1
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    • pp.1-11
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    • 2012
  • We identified cdf based on screening of the Arabidopsis cDNA library for functional suppressors of the AtBI-1 (a gene described to suppress the cell death induced by Bax gene expression in yeast). The cdf was located on Chr. V and was composed of 5 exons and 4 introns. It encodes a protein of 258 amino acid residues with a molecular weight of 28.8 kDa. The protein has 3 transmembrane domains in the C-terminal region. The cdf has one homologue, named cdf2, which was found in Arabidopsis. Like cdf, cdf2 also induced growth defect in yeast. The effect of the cell growth defect factor was somewhat lower than Bax. cdf could arrest the growth of yeast. Its localization to the nucleus was essential for the suppression of yeast cell proliferation. Morphological abnormality of intracellular network, which is a hallmark of AtBI-1, was attenuated by expression of cdf.

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Effect of IPTG Induction on Production of ${\beta}$-Galactosidase-PreS2 Fusion Protein in Recombinant Escherichia coli

  • Nam, Soo-Man;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.274-280
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    • 1991
  • Effects of IPTG induction on cell growth and production of ${\beta}$-galactosidase-preS2 fusion protein (${\beta}$gal-preS2) were studied in a defined medium using a recombinant Escherichia coli JM109/pCMHB30. IPTG was added (0.2 mM) to induce the cloned-gene expression in the early-, mid-, and late-log growth phases. The most serious decreases in growth rate and plasmid stability were observed for the induction in the early-log growth phase. The expression level of ${\beta}$gal-preS2 attained by the induction in the mid-log phase was about 0.51 mg fusion protein/mg total cellular protein, which was 2- and 5-fold improvement over the levels obtained with the inductions in the early- and late-log phases. Formation of acidic byproducts including acetate and pyruvate showed different profiles during the fermentation period for each cases of induction; pyruvate was the major byproduct for the induction in the early-log phase while acetate production became more significant for the cases of inductions in the mid- and late-log phases.

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Neuronal Differentiation of PC12 Cells Cultured on Growth Factor-Loaded Nanoparticles Coated on PLGA Microspheres

  • Park, Keun-Hong;Kim, Hye-Min;Na, Kun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1490-1495
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    • 2009
  • The development of nanotechnology has penetrated the fields of biology and medicine, resulting in remarkable applications for tissue regeneration. In order to apply this technology to tissue engineering, we have developed nano-scaled 3D scaffolds consisting of growth factor-loaded heparin/poly(l-lysine) nanoparticles (NPs) attached to the surface of polymeric micro spheres via polyionic complex methods. Growth factor-loaded NPs were simply produced as polyelectrolyte complexes with diameters of 100-200 nm. They were then coated onto positively charged poly(lactic-co-glycolic acid) (PLGA) pretreated with polyethyleneimine to enable cell adhesion, proliferation, and stimulation of neurite outgrowth. Propidium iodide staining and $\beta$-tubulin analysis revealed that neuronal PC12 cells proliferated extensively, expressed significant amounts of b-tubulin, and showed well-structured neurite outgrowth on polymeric microspheres by stimulation with growth factors. These results suggest that cellular adhesion and biological functionality on prepared PLGA microspheres enabled terminal differentiation of neuronal cells.

Isolation and Characterization of a Thermophilic Bacillus sp. producing a Thermostable $\alpha$-glucosidase (내열성$\alpha$-glucosidase를 생산하는 호열성 Bacillus sp. 균주의 분리 및 특성)

  • 이용억
    • Journal of Life Science
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    • v.8 no.4
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    • pp.387-394
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    • 1998
  • A thermophilic bacterium (strain DG0303) producing a thermostable $\alpha$-glucosidase was isolated from manure and identified as Bacillus sp. Strain DG0303 produced high level of $\alpha$-glucosidase compared with other thermophilic Bacillus strains. The cellular protein patterns were also compared with other Bacillus strains by sodium dodecyl sulfatepolyacrylamide gel electrophoresis(SDS-PAGE). On the basis of 16S rDNA analysis the Bacillus sp. DG0303 was found to be a member of Bacillus rDNA group 5. The optimum temperature for growth was 65$\circ$C and no growth was obtained at 40$\circ$C or 75$\circ$C. The optimum pH for growth was 5.5 to 8.5. $\alpha$-glucosidase activity was produced during growth and most activity was detected in the culture supernatant. The $\alpha$-glucosidase production was constitutive in the absence of carbohydrates. High level of enzyme activity was detected when the culture was grown on medium containing starch. Addition of glucose resulted in the repression of the $\alpha$-glucosidase production. The optimum pH and tempoerature for enzyme activity were pH 5.0 and 65$\circ$C, respectively. When analyzed by zymogram, the culture supernatant showed a single $\alpha$-glucosidase band with a molecular weight of approximately 60,000.

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A Basic Helix-Loop-Helix Transcription Factor Regulates Cell Elongation and Seed Germination

  • Kim, Jin-A;Yun, Ju;Lee, Minsun;Kim, Youn-Sung;Woo, Jae-Chang;Park, Chung-Mo
    • Molecules and Cells
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    • v.19 no.3
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    • pp.334-341
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    • 2005
  • Plants are sessile and rely on a wide variety of growth hormones to adjust growth and development in response to internal and external stimuli. We have identified a gene, designated NAN, encoding a basic helix-loop-helix (bHLH) transcription factor that regulates cell elongation and seed germination in plants. NAN has an HLH motif in its C-terminal region but does not have any other discernible homologies to bHLH proteins. A bipartite nuclear localization signal is located close to the HLH motif. An Arabidopsis mutant, nan-1D, in which NAN is activated by the insertion of the 35S enhancer, exhibits growth retardation with short hypocotyls and curled leaves. It is also characterized by reduced seed germination and apical hook formation, symptomatic of GA deficiency or disrupted GA signaling. The phenotypic effects of nan-1D were increased by treatment with paclobutrazol (PAC), an inhibitor of gibberellic acid (GA) biosynthesis. NAN is constitutively expressed throughout the life cycle. Our observations indicate that NAN has a housekeeping role in plant growth and development, particularly in seed germination and cell elongation, and that it may modulate GA signaling.

Inhibition of Cell Proliferation and Migration by miR-509-3p That Targets CDK2, Rac1, and PIK3C2A

  • Yoon, Sena;Han, Eunji;Choi, Young-Chul;Kee, Honghwan;Jeong, Yongsu;Yoon, Jaeseung;Baek, Kwanghee
    • Molecules and Cells
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    • v.37 no.4
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    • pp.314-321
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    • 2014
  • CDK2 is a key regulator of cell cycle progression. In this study, we screened for miRNAs targeting CDK2 using a luciferase-3'-untranslated region reporter assay. Among 11 hit miRNAs, miR-509-3p reduced CDK2 protein levels and significantly inhibited cancer cell growth. Microarray, Western blotting, and luciferase reporter analyses revealed additional targets of miR-509-3p, including Rac1 and PIK3C2A. Overexpression of miR-509-3p induced G1 cell-cycle arrest and inhibited colony formation and migration. RNAi experiments indicated that the growth-inhibitory effects of miR-509-3p may occur through down-regulation of CDK2, Rac1, and PIK3C2A. Targeting of multiple growth regulatory genes by miR-509-3p may contribute to effective anti-cancer therapy.

Cholesterol Uptake by Lactobacillus acidophilus: Its Fate and Factors Influencing the Uptake (Lactobacillus acidophilus에 의한 콜레스테롤의 흡착)

  • Noh, Dong-Ouk
    • Journal of Dairy Science and Biotechnology
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    • v.14 no.2
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    • pp.195-206
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    • 1996
  • Cholesterol assimilated by Lactobacillus acidophilus ATCC 43121 was not metabolically degraded in that most of it was recovered with the cells. Cells grown in the presence of cholesterol micelles and bile salts were more resistant to Iysis by sonication than those grown in their absence, suggesting a possible alteration of cellular membranes. Cholesterol assimilation occurred during growth at pH 6.0, the amount of which was more than that by cells grown without pH control. Cholesterol assimilated by cells was recovered in the membrane fractions of cells both grown at pH 6.0 and without pH control. The effect of unsaturated fatty acids on cholesterol assimilation was not clear, since there was no significant (P> 0.05) difference in the amount taken up from micelles prepared using L-${\alpha}$-phosphatidylcholine, dioleoyl or L-${\alpha}$-phosphatidylcholine, distearoyl. Without Tween 80, little, if any, cell growth or cholesterol uptake was observed. In the presence of 0.05% Tween 80, cholesterol uptake increased dramatically as did growth. However, as the amount of Tween 80 increased beyond 0.05%, cholesterol uptake decreased while the amount of growth remained the same.

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Comparative Analysis of Growth-Phase-Dependent Gene Expression in Virulent and Avirulent Streptococcus pneumoniae Using a High-Density DNA Microarray

  • Ko, Kwan Soo;Park, Sulhee;Oh, Won Sup;Suh, Ji-Yoeun;Oh, TaeJeong;Ahn, Sungwhan;Chun, Jongsik;Song, Jae-Hoon
    • Molecules and Cells
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    • v.21 no.1
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    • pp.82-88
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    • 2006
  • The global pattern of growth-dependent gene expression in Streptococcus pneumoniae strains was evaluated using a high-density DNA microarray. Total RNAs obtained from an avirulent S. pneumoniae strain R6 and a virulent strain AMC96-6 were used to compare the expression patterns at seven time points (2.5, 3.5, 4.5, 5.5, 6.0, 6.5, and 8.0 h). The expression profile of strain R6 changed between log and stationary growth (the Log-Stat switch). There were clear differences between the growth-dependent gene expression profiles of the virulent and avirulent pneumococcal strains in 367 of 1,112 genes. Transcripts of genes associated with bacterial competence and capsular polysaccharide formation, as well as clpP and cbpA, were higher in the virulent strain. Our data suggest that late log or early stationary phase may be the most virulent phase of S. pneumoniae.