• Title/Summary/Keyword: 4E-BP

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Isolation and Characterization of Eukaryotic Translation Initiation Factor 5A (eIF-5A) from Potato (감자로부터 Eukaryotic Translation Initiation Factor 5A (elF-5A) 유전자의 동정 및 발현 분석)

  • 인준교;신동호;최관삼;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.283-287
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    • 2001
  • Differential display based on PCR was employed to identify genes expressed during tuber-developing stage of potato (Solanum tuberosum L. cv. Irish Cobbler). An eukaryotic initiation factor 5A (eIF-5A) clone isolated from a cDNA library constructed with developing micro-tuber using a probe of PCR fragment. We isolated three positive clones and ore of them contained open reading frame. This clone revealed high sequence similarity to tomato eIF 5A cDNA. At the DNA level, there is 94.8% identity with the tomato eIF-5A4, whereas at the protein level there is a high identity with 97.5%. The potato eIF 5A clone is 716 bp in length and contains a single open reading frame from 57 to 539 bp, a 56 bp 5'-untranslated region and a 177 bp 3'-untranslated region. The deduced protein composed of 160 amino acid residues, with a predicted molecular mass of 17.4 kD and an estimated pl of 5.5. The sequence of 12 (STSKTGKHGHAK) amino acids among eIF-5A proteins is perfectly conserved from yeast to human. That sequence in potato eIF-5A protein is also conserved at position 46 to 57 amino acid. This region embeds the post-translational modification site of the lysine residue (at the seventh K) to hypusine that is crucial to eIF-5A activity. The northern blot analysis of eIF5A has shown abundant expression, mainly in flower organs (stamen, ovary, petal, sepal), fruit and stolen.

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Ceramide Induces Apoptosis and Growth Arrest of Human Glioblastoma Cells by Inhibiting Akt Signaling Pathways

  • Lee, Eun-Chang;Lee, Young-Seok;Park, Na-Hee;So, Kwang-Sup;Chun, Young-Jin;Kim, Mie-Young
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.21-26
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    • 2011
  • Ceramide is an important lipid mediator of extracellular signals that control various cellular functions, including apoptosis. In this study, we showed that ceramide induced apoptosis in U373MG human glioblastoma cells associated with G1 cell cycle arrest. Treatment of cells with ceramide increased proapoptotic Bax expression and inhibited the expression of antiapoptotic Bcl-2 and Bcl-xL Ceramide also downregulated cyclin E, cyclin D1, cdk 2, and cdk4 which are involved in regulating cell cycle. In addition, ceramide suppressed phosphorylation of Akt, Bad, p70 S6 kinase, and 4E-BP1, suggesting the involvement of Akt/mTOR signaling pathway. Additionally, okadaic acid, an inhibitor of protein phosphatase 2A, partially blocked the ceramide mediated inhibition of phosphorylation of Akt and 4E-BP1. These results suggest that ceramide induces apoptosis in U373MG glioblastoma cells by regulating multiple signaling pathways that involve cell cycle arrest associated with Akt signaling pathway.

Molecular Cloning of a Cellulase Gene from Abalone Haliotis discus hannai and Its Expression in E coli

  • Park, Eun-Mi;Han, Yun-Hee;Park, In-Suk;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Kim, Young-Ok
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.2
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    • pp.108-112
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    • 2007
  • A cellulase (endo-${\beta}$-1,4-D-glucanase(E.C.3.2.1.4)) was isolated from the hepatopancreas of abalone Haliotis discus hannai by EST analysis. The abalone cellulase named HdEG compassed 1977 bp, including 195 bp in the 5'untranslated region, 1680 bp in the open reading frame which encodes 560 amino acid residues, and 92 bp in the 3'-untranslated region. The C-terminal region of the HdEG showed 44-52% identity to the catalytic domains of glycoside hydrolase family 9 (GHF9)-cellulases from arthropods and bacteria. The recombinant cellulase, pEHdEG was produced in E. coli with being fused with C-terminal His-tag. The expressed protein showed a single band (~62 kDa) on Western blotting which was consistent with the value (61,878 Da) calculated from the DNA sequence.

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The Environmental Change of Korea based on the Isopollen Map during the Holocene

  • Yoon, Soon-Ock
    • The Korean Journal of Quaternary Research
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    • v.22 no.2
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    • pp.6-11
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    • 2008
  • Vegetation change reconstructed by pollen analysis is effective to clarify natural conditions such as climate and soil as well as intensity of human activity. Pollen analysis in Korea is difficult to obtain peaty soil sedimented by low relief geomorphollogically and formation age is usually confined to obtain information during young Holocene as well as little absolute age data. Isopollen map was constructed in order to analyze the change of vegetation environment time-spatially during Holocene based on the 30 data with age dated from 78 results from pollen analysis in Korea. The indicatives for vegetation environment were the main trees in Korea such as Alnus, Pinus, Quercus and AP/NAP during the periods of 6,000 y.BP, 4,000 y.BP, 3,000 y.BP, 2,000 y.BP, 1,000 y.BP. As a result, the regional time-spatial patterns of vegetation distribution appeared clearly on the isopollen map. The dominant vegetation stage was repeated in the different pattern e.g. the dominance between Alnus and Quercus at West Coast and between Pinus and Quercus at East Coast competitively.

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4-(N-Methyl-N-nitrosamino)-1(3-pyridyl)-1-butanone(NNK) Restored the Cap-dependent Protein Translation Blocked by Rapamycin

  • Kim Jun-Sung;Park Jin Hong;Park Sung-Jin;Kim Hyun Woo;Hua Jin;Cho Hyun Sun;Hwang Soon Kyung;Chang Seung Hee;Tehrani Arash Minai;Cho Myung Haing
    • Toxicological Research
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    • v.21 no.4
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    • pp.347-353
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    • 2005
  • Eukaryotic initiation factor 4E (elF4E) is a key element for cap-dependent protein translation controlled by affinity between elF4E and 4E-binding protein 1 (4E-BP1). Rapamycin can also affect protein translation by regulating 4E-BP1 phosphorylation. Tobacco-specific nitrosamine, 4(N-methyl-N-nitrosamino )-1-(3-pyridyl)-1-butanone (NNK) is a strong lung carcinogen, but its precise lung cancer induction mechanism remains unknown. Relative roles of cap-dependent and -independent protein translation in terms of NNK-induced lung carcinogenesis were elucidated using normal human bronchial epithelial cells. NNK concentrations applied in this study did not decrease cell viability. Addition of NNK restored rapamycin-induced decrease of protein synthesis and rapamycin-induced phosphorylation of 4E-BP1, and increased expression levels of mTOR, ERK1/2, p70S6K, and Raf-1 in a concentration-dependent manner. NNK also caused perturbation of normal cell cycle progression. Taken together, NNK might cause toxicity through the combination of restoration of 4E-BP1 phosphorylation and increase of elF4E as well as mTOR protein expression, interruption of Raf1/ERK as well as the cyclin G-associated p53 network. Our data could be applied towards elucidation of the molecular basis for lung cancer treatment.

Estimating BP Decoding Performance of Moderate-Length Irregular LDPC Codes with Sphere Bounds

  • Chung, Kyu-Hyuk
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.35 no.7C
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    • pp.594-597
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    • 2010
  • This paper estimates belief-propagation (BP) decoding performance of moderate-length irregular low-density parity-check (LDPC) codes with sphere bounds. We note that for moderate-length($10^3{\leq}N{\leq}4\times10^3$) irregular LDPC codes, BP decoding performance, which is much worse than maximum likelihood (ML) decoding performance, is well matched with one of loose upper bounds, i.e., sphere bounds. We introduce the sphere bounding technique for particular codes, not average bounds. The sphere bounding estimation technique is validated by simulation results. It is also shown that sphere bounds and BP decoding performance of irregular LDPC codes are very close at bit-error-rates (BERs) $P_b$ of practical importance($10^{-5}{\leq}P_b{\leq}10^{-4}$).

The Effect of Angelica gigas Nakai Extract and Bacillus Polyfermenticus KJS-2 on Atopic Dermatitis induced by DNCB in mice (DNCB로 유도된 아토피 피부염에 대한 당귀 추출물과 Bacillus polyfermenticus KJS-2의 효과)

  • Ryu, Deok-Hyun;Oh, Sa-Rang;Jung, Tae-Sung;Ryu, Deok-Seon
    • The Journal of Korean Medicine
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    • v.38 no.3
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    • pp.30-42
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    • 2017
  • Objectives: The purpose of this study was to investigate the effects of Angelica gigas Nakai extract(AGNE) and Bacillus polyfermenticus KJS-2 (BP2) on atopic dermatitis (AD) induced by 2, 4-dinitrochlorobenzene (DNCB) in mice. Methods: In the experiment, we divided mice into four groups: a control group, a DNCB group, an AGNE group, and an AGNE+BP2 group. Then we examined the changes in scratching frequency, clinical aspects on dorsum skin, immunoglobulin (IgE), cytokines ($TNF-{\alpha}$, IL-6) and expression of COX-2. Resutls: From the experiment, the scratching frequency was significantly dropped in AGNE group and AGNE+BP2 group. Clinical observations of dorsum skin, there were a severe keratotic lesion and drop of dead skin cell in DNCB group, but symptoms of AD were decreased 39.6% in AGNE group and 49.6% in AGNE+BP2 group during 3 weeks. IgE, $TNF-{\alpha}$ and IL-6 were decreased significantly in both AGNE and AGNE+BP2 group. Expression of COX-2 was also decreased significantly in both groups. Conclusions: In conclusion, these data suggest that AGNE can decrease symptoms of AD and BP2 makes AGNE more effective. So AGNE can be useful herbal therapy for AD.

Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.211-218
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    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

Production of the recombinant fibronectin-binding protein of Staphylococcus aureus (Staphylococcus aureus의 재조합 fibronectin-binding protein의 생산)

  • kim, Doo;Cheong, Cha-ryong;Park, Hee-myong;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.37 no.4
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    • pp.875-882
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    • 1997
  • To produce the recombinant fibronectin-binding protein(FnBP) for development of subunit vaccine against Staphylococcus aureus. The fnbp gene was amplified from the chromosomal DNA of S aureus KNU 196 strain using the polymerase chain reaction, and cloned into pGEX-4T-2. Then, the recombinant FnBP fused with glutathione-S-transferase was produced in E coli, purified by affinity chromatography, and identified its antigenicity and immunogenicity by Western blot. The recombinant FnBP produced in this study is considered to have the same property of native FnBP purified from S aureus, and is expected to be useful as a candidate for S aureus subunit vaccine.

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The fruit of Acanthopanax senticosus Harms improves arterial stiffness and blood pressure: a randomized, placebo-controlled trial

  • Oh, Eunkyoung;Kim, Youjin;Park, Soo-yeon;Lim, Yeni;Shin, Ji-yoon;Kim, Ji Yeon;Kim, Ji-Hyun;Rhee, Moo-Yong;Kwon, Oran
    • Nutrition Research and Practice
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    • v.14 no.4
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    • pp.322-333
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    • 2020
  • BACKGROUND/OBJECTIVES: Arterial stiffness and endothelial dysfunction are 2 of the independent predictors for cardiovascular disease, while Acanthopanax senticosus Harms (ASH) is a traditional medicinal plant that can improve cardiovascular health. This study aimed to investigate the efficacy of the fruit of ASH on vascular function in apparently healthy subjects. SUBJECTS/METHODS: A 12-week, randomized, double-blind, placebo-controlled design, consisting of healthy adults with at least 2 of the following 3 conditions: borderline high blood pressure (BP; 120 mmHg ≤ systolic BP ≤ 160 mmHg or 80 mmHg ≤ diastolic BP ≤ 100 mmHg), smoking (≥10 cigarettes/day), and borderline blood lipid levels (220 ≤ total cholesterol ≤ 240, 130 ≤ low density lipoprotein cholesterol ≤ 165, or 150 ≤ triglyceride ≤ 220 mg/dL). Randomly assigned 76 subjects who received a placebo or 2 doses of ASH fruit (low, 500 mg/day; high, 1,000 mg/day) completed the intervention. Brachial-ankle pulse wave velocity (baPWV), flow-mediated dilation, carotid intima-media thickness, and BP were measured both at baseline and following the 12-week intervention. Endothelial nitric oxide synthase (eNOS) phosphorylation was assessed by western blotting. RESULTS: Compared with the placebo group, the low-dose group showed more significant changes after the 12-week intervention period in terms of systolic BP (0.1 vs. -7.7 mmHg; P = 0.044), baPWV (31.3 vs. -98.7 cm/s; P = 0.007), and the ratio of phospho-eNOS/eNOS (0.8 vs. 1.22; P = 0.037). CONCLUSIONS: These results suggest that ASH fruit extract at 500 mg/day has the potential to improve BP and arterial stiffness via endothelial eNOS activation in healthy adults with smoking and the tendency of having elevated BP or blood lipid parameters.