• Title/Summary/Keyword: Regulation of Protein Synthesis

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Association of Anti-apoptotic Mechanism Due to House Dust Mite in Neutrophils with Protein Synthesis and Bad

  • Kim, In Sik;Lee, Ji-Sook
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.211-214
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    • 2016
  • House dust mite is an essential allergen in the pathogenesis of allergic diseases. Abnormal regulation of neutrophil apoptosis is an important pathogenic process in allergic diseases. In the present study, we investigated the effects of house dust mites on spontaneous apoptosis of neutrophils and its associated mechanisms. Extract of Dermatophagoides pteronissinus (DP) inhibited neutrophil apoptosis in a time-dependent manner. Cycloheximide (CHX), an inhibitor of translation, increased apoptosis of DP-treated neutrophils as well as control cells. The pro-apoptotic effect of CHX was blocked by DP in neutrophils. In addition, DP increased the phosphorylation of Bad in a time-dependent manner, indicating that it exerted an inhibitory effect on the function of Bad. These results suggest that DP has anti-apoptotic effects of neutrophils and may regulate protein synthesis and activation of Bad. Moreover, these findings may shed light on elucidation of allergy pathogenesis due to house dust mites.

REGULATION OF PROENKEPHALIN GENE EXPRESSION AND MET-ENKEPHALIN SECRETION IN BOVINE ADRENAL MEDULLARY CHROMAFFIN CELLS AND C6 RAT GLIOMA CELLS

  • Suh, Hong-Won
    • Toxicological Research
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    • v.9 no.2
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    • pp.195-206
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    • 1993
  • The expression of proenkephalin (proENK) mRNA and Met-enkephalin (ME) secretion in C6 rat glioma cells and bovine adrenal medullary chromaffin (BAMC) cells were elucidated in the present study. The levels of proENK mRNA and ME secreted into the media in BAMC cells were measured in the presence of cycloheximide and 12-tetrade-canoylphorbol-13-acetate (TPA). Cycloheximide (20 nM) abolished the induction of proENK mRNA expression, protein synthesis and ME secretion by TPA (1nM), indicating that de novo protein synthesis was necessary for proENK gene expression and ME secretion.

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Regulation of Ferritin Synthesis by Iron-responsive Element in 5'-Untranslated Region (5'-Untranslated Region에 존재하는 Iron Responsive Element에 의한 Ferritin 합성조절)

  • Chung, In-Sik;Lee, Jung-Lim;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.41 no.3
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    • pp.224-227
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    • 1998
  • The expression of ferritin involved in iron metabolism is regulated at the translational level by the interaction of iron regulatory protein with iron-responsive element(IRE) in the 5'-untranslated region of ferritin transcript. To identify the role of structural element utilized for translational regulation of ferritin, we studied the effects of mutations in the ferritin IRE by measuring IRP binding activity and translational activity. Our data suggest that the cytosine at bulged position of IRE within ferritin is important for the formation of RNA secondary structure involved in translational regulation.

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Neural Tissue-Specific Epidermal Growth Factor (EGF)-like Domain Containing Protein, NELL2, Plays on Important Role in the Control Regulation of Puberty Onset in the Female Rat Hypothalamus

  • Ha, Chang-Man;Kang, Hae-Mook;Lee, Byung-Ju
    • Animal cells and systems
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    • v.4 no.4
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    • pp.367-373
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    • 2000
  • In the present study we determined if NELL2, a neural tissue-specific protein containing 6 epidermal growth factor (EGF)-like repeat domains, plays an important role in the regulation of puberty initiation in the rat hypothalamus. We origin811y found that NELL2 is a new estrogen-responsive gene in hypothalami derived from estrogen-sterilized and control rats using a PCR differential display. In the 40-day-old female rat hypothalamus, NELL2 was up-regulated by neonatal estrogen treatment. In situ hybridization histochemistry showed that NELL2 is very abundant in the ventromedial hypothalamic nucleus that is responsible for the control of sex behavior. NELL2 mRNA level in the medial basal hypothalamus showed a dramatic increase before female puberty onset, which suggests that NELL2 may be involved in the process regulating female puberty onset. We attemped to block NELL2 synthesis with intracerebroventricular injection of an antisense oligodeoxynucleotide (ODN) to the NELL2 mRNA, and examined its effect on the puberty onset of the female rat. The antisense ODN significantly delayed puberty initiation determined by vaginal opening. In summary, NELL2 may play an important role in the regulation of female puberty onset.

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Regulation of Lipoprotein Lipase by Fasting in Epididymal and Mesenteric Adipocytes of Rats

  • Lee, Jae Joon;Chung, Chung Soo;Lee, Myung Yul
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.5
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    • pp.715-722
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    • 2008
  • There are marked variations in the activity of lipoprotein lipase (LPL) among adipose depots. The aim of this study was to compare the mechanisms of 24 h of fasting on LPL regulation between epididymal (EPI) adipocytes and mesenteric (MES) adipocytes in rats. 1-Day fasting consistently decreased activities of heparin-releasable LPL, total extractable LPL and cellular LPL markedly in both EPI and MES fat pads. LPL activity in MES fat pads was relatively lower than in the EPI fat pads. Consistent with data on LPL activity, the levels of expression of LPL mRNA in both nutritional states were lower in MES than EPI adipose tissue and isolated adipocytes. The decreased LPL activity after 1 day of fasting in MES adipocytes was explained mainly by a 50% decrease in the relative abundance of LPL mRNA level and a parallel 50% decrease in relative rate of LPL synthesis. In contrast, fasting of 1 day in EPI adipocytes decreased total LPL activity by 47% but did not affect LPL mRNA level or relative rate of LPL synthesis. A decrease in overall protein synthesis contributed to the decreased LPL activity after 1 day fasting both in EPI and MES adipocytes. In MES adipocytes the decrease in LPL activity, LPL mRNA and LPL synthesis were comparable, but in EPI adipocytes the changes in LPL activity were substantially larger than the changes in LPL mRNA level and LPL synthesis. Therefore, fasting decreased fat cell size, LPL activity, LPL mRNA level and relative rate of LPL synthesis in rats, and these effects were more marked in the MES adipocytes. These results clearly demonstrate the regional variations in the metabolic response of adipose tissue and LPL functions to fasting.

Regulation of adductor muscle growth by the IGF-1/AKT pathway in the triploid Pacific oyster, Crassostrea gigas

  • Kim, Eun-Young;Choi, Youn Hee
    • Fisheries and Aquatic Sciences
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    • v.22 no.9
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    • pp.19.1-19.10
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    • 2019
  • We investigated the insulin-like growth factor 1 (IGF-1)/AKT signaling pathway involved in muscle formation, growth, and movement in the adductor muscle of triploid Pacific oyster, Crassostrea gigas. Large and small triploid oysters (LTs and STs) cultured under identical conditions were screened, and the signaling pathways of individuals with superior growth were compared and analyzed. mRNA and protein expression levels of actin, troponin, tropomyosin, and myosin, proteins important in muscle formation, were higher in LTs compared with STs. Expression levels of IGF-1, IGF binding protein (IGFBP), and IGFBP complex acid-labile subunit were also higher in LTs compared with STs. Phosphorylation of the IGF receptor as well as that of AKT was high in LTs. In addition, the expression of phosphomammalian target of rapamycin and phospho-glycogen synthase kinase $3{\beta}$ was increased and the expression of Forkhead box O3 was decreased in LTs. Therefore, we suggested that the IGF-1/AKT signaling pathway affects the formation, growth, and movement of the adductor muscle in triploid oysters.

Regulation of Compaction by Synthesis and Phosphorylation of Protein in Preimplantation Mouse Embryo (생쥐 배아에서 단백질 합성과 인산화에 의한 밀집현상의 조절)

  • 이동률;이정은;윤현수;노성일;김문규
    • Development and Reproduction
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    • v.3 no.1
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    • pp.75-85
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    • 1999
  • To investigate the origin and action mechanism of cytoplasmic factors as regulators of morphogenesis, the embryonic development, RNA synthesis and protein phosphorylation were examined in reconstituted embryos. A half of 1-cell mouse embryo with both pronuclei was electrofused with the enucleated cytoplasm of 1- or 2-cell embryos which were cultured for 24 hrs from post 20 hrs hCG in CZB with or without cycloheximide (CHX, an inhibitor of protein synthesis; P+P-CHX group), genistein (Gen, an inhibitor of tyrosine protein kinase; P+2-Gen group) and 6-dimethylaminopurine (6-DMAP, an inhibitor of serine-threonine protein kinase; P+2-DMAP group), and co-cultured with Vero cells for 5 days. And their development, cell numbers at compaction, [5, 6-$^3$H]-uridine incorporation into RNA and the pattern of protein phosphorylation after labeling of [$^{32}$ P] orthophosphate were compared with that of the reconstituted embryos such as P+2 or P+P (control group). Embryonic development and the time of RNA synthesis in P+P-CHX were similar to those in P+P. But the time and the cell stages of embryonic compaction in P+P-CHX were similar to those in P+2. The compaction was initiated at 4-cell in P+2 and P+2-Gen, but at 8-cell in P+P and P+2-DMAP. On a two-dimensional gel electrophoresis, phosphorylation of 80KD and 110KD proteins were inhibited after 3 hrs of reconstruction in the embryo of P+2-DMAP when compared with that of P+2 and P+2-Gen. These results suggest that protein synthesis between 1- and 2-cell stage affects the timing of embryonic genome activation, and that cytoplasmic factors derived from oocyte or their modification regulates the time schedule of embryonic compaction in mouse. Also, serine-threonine protein kinase has an important role on the regulation of compaction.

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Cloning and Regulation of Schizosaccharomyces pombe Gene Encoding Ribosomal Protein S20

  • Lee, Yoon-Jong;Kim, Kyunghoon;Park, Eun-Hee;Ahn, Ki-Sup;Kim, Daemyung;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.31-36
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    • 2001
  • A cDNA clone encoding the ribosomal protein S20 has been isolated from the Schizosaccharomyces pombe cDNA library by colony hybridization. The insert contained in the original plasmid pYJ10 was transferred intro shuttle vector pRS316 generate plasmid pYJll. The dDNA insert of plasmid pYJll, contains 484 nucleotides and encodes a protein of 118 amino acids with a calculated mass of 13,544 daltons. The deduced amino acid sequence of S. pombe ribosomal protein S20 is very homologous with fruit fly, rat, and budding yeast counterparts. It is also homologous with Xenopus S22 ribosomal protein. S. pombe ribosomal protein S20 appears to be relatively hydruphobic except the C-terminal region. The 728 bp upstream region of the S20 gene was amplified from chromosomal DNA and transferred into the BamHI/EcoRI site of the promoterles $\beta$-galactosidase gene of the vector YEp357R, which resulted in fusion plasmid pYS20. The synthesis of $\beta$-galactosidase from the fusion plasmid appeared to be the highest in the mid-exponential phase. The S. pombe cells with the fusion plasmid grown at 35$\^{C}$ gave lower $\beta$-galactosidase activity than the cells grown at 30$\^{C}$. Computer analysis showed the consensus sequence CAGTCACA in the upstream regions of various ribosomal protein genes in S. pombe, which would be involved in the coordinated expression of small ribosomal proteins.

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Hormonal Regulation of Insulin-Like Growth Factor Binding Protein Secretion by a Bovine Mammary Epithelial Cell Line

  • Kim, W.Y.;Chow, J.C.;Hanigan, M.D.;Calvert, C.C.;Ha, J.K.;Baldwin, R.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.2
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    • pp.233-239
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    • 1997
  • A mammary epithelial cell line (MAC-T) established as a model for lactation was utilized to identify and characterize effects of various hormones upon insulin-like growth factor binding protein secretion. Ligand and immunoblot analyses of conditioned media indicated that insulin-like growth factor binding protein-2 was secreted by MAC-T cells. Insulin-like growth factor-I stimulated insulin-like growth factor binding protein-2 secretion in a dose-dependent manner, but prolactin and bovine somatotropin did not alter insulin-like growth factor binding protein-2 secretion. Insulin increased and cortisol decreased insulin-like growth factor binding protein-2 secretion. Effects of insulin-like growth factor-I on insulin-like growth factor binding protein-2 secretion support previous studies using primary cultures of bovine mammary cells and bovine fibroblasts. Effects of cortisol and insulin on insulin-like growth factor binding protein-2 secretion may be explained by changes in protein synthesis. In addition, supraphysiological doses of insulin can cross-react with the insulin-like growth factor-I receptor and stimulate insulin-like growth factor binding protein-2 secretion. MAC-T cells provide a model system to study mechanisms that regulate local insulin-like growth factor-I bioactivity.

Effect of ganglioside GD3 synthase gene expression on VSMC proliferation via ERK1/2 pathway, cell cycle progression and MMP-9 expression

  • Lee, Young-Choon;Kim, Cheorl-Ho
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2004.10a
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    • pp.81-90
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    • 2004
  • Sialic acid containing glycosphingolipids (gangliosides) have been implicated in the regulation of various biological phenomena such as atherosclerosis. Recent report suggeststhat exogenously supplied disialoganglioside (GD3) serves a dual role in vascular smooth muscle cells (VSMC) proliferation and apoptosis. However, the role of the GD3 synthase gene in VSMC responses has not yet been elucidated. To determine whether a ganglioside is able to modulate VSMC growth. the effect of overexpression of the GD3 synthase gene on DNA synthesis was examined. The results show that the overexpression of this gene has a potent inhibitory effect on DNA synthesis and ERK phosphorylation in cultured VSMC in the presence of PDGF. The suppression of the GD3 synthase gene was correlated with the down-regulation of cyclinE/CDK2. the up-regulation of the CDK inhibitor p21 and blocking of the p27 inhibition,whereas up-regulation of p53 as the result of GD3 synthase gene expression was not observed. Consistently, blockade of GD3 function with anti-GD3 antibody reversed VSMC proliferation and cell cycle proteins. The expression of the CD3 synthase gene also led to the inhibition of TNF--induced matrix metalloproteinase-9 (MMP-9) expression in VSMC as determined by zymography and immunoblot. Furthermore, GD3 synthase gene expression strongly decreased MMP-9 promoteractivlty in response to TNF-. This inhibition was characterized by the down-regulation of MMP-9,which was Iranscriptionally regulated at NF-B and activation protein-1 (AP-1) sites in the MMP-9promoter Finally, the overexpression of MMP-9 in GD3 synthase transfectant cells rescued VSMC proliferation. However MMP-2 overexpression was not affected the cell proliferation. These findings suggest that the fl13 synthase gene represents a physiological modulator of VSMC responses that may contribute to plaque instability in atherosclerosis.

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