• Title/Summary/Keyword: Sterol biosynthesis

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Methylation of CpG Islands in the Rat 7-dehydrocholesterol Reductase Promoter Suppresses Transcriptional Activation

  • Kim, Jai-Hyun;Hwang, Eun-Ha;Park, Hye-Jung;Paik, Young-Ki;Shim, Yhong-Hee
    • Molecules and Cells
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    • v.19 no.2
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    • pp.279-282
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    • 2005
  • In mammals, 7-dehydrocholesterol reductase (Dhcr7) is the terminal enzyme in cholesterol biosynthesis. We previously reported that the Dhcr7 proximal promoter (-179 to +1), which contains CpG islands, is responsible for sterol-mediated expression of the rat gene. In the present study, we examined whether methylation of this region affects the transcriptional activity of the Dhcr7 gene. In vitro DNA methylation of the Dhcr7 promoter and luciferase-reporter assays showed that DNA methylation of the CpG islands suppressed transcription. Furthermore, treatment of the methylated Dhcr7 promoter with the demethylating agent, 5-aza-2'-deoxycytidine (5-Aza-CdR), reversed the suppression of promoter activity. These results indicate that methylation of the CpG islands is an important transcriptional regulatory mechanism in the Dhcr7 promoter.

Insecticidal Efficacy of Porteresia coarctata (Roxb.) on Bio-chemical Alteration of Spodoptera litura (Fab.)

  • Bhattacharyya Atanu;Goswami Arunava;Dihidar Shankar;Adhikari Sujit;Banerjee Sudip Kumar;Talapatra Soumendra Nath;Barik Bikas Ranjan;Bhattacharya Subir Kumar;Chandra Asit Kumar
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.1
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    • pp.9-13
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    • 2007
  • Hexane extract of Porteresia coarctata (Roxb.) exhibits a toxic effect on the tissues of Spodoptera litura (F) while fed at the dose of 1000 and 2000 ppm thoroughly mixing with castor leaves (Ricinus communis L) after dissolving in DMSO at late fourth instar whereas only DMSO treated castor leaves were fed to control group. The larvae were put to rear at $28^{\circ}C{\pm}1^{\circ}C$, $76{\pm}4%$ R.H. under 12 L + 12 D photoperiodic regime. In test group insects substantial reduction of protein and DNA content was marked in fat body and midgut tissues compared to DMSO treated control group. The significant biochemical alterations in the midgut tissues and fat body of test group insects indicate the insecticidal property of the said plant extract that could be tested in facilitating the phenomenal stride in Integrated Pest Management.

Effect of Oral Administration of Egg Yolk on Cholesterol Metabolism in Rats (난황 경구투여가 랫드의 콜레스테롤 대사에 미치는 영향)

  • Bang, Han-Tae;Hwangbo, Jong;Park, Sang-Oh;Park, Byung-Sung
    • Journal of the Korean Applied Science and Technology
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    • v.31 no.2
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    • pp.255-264
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    • 2014
  • This study was performed to elucidate the biochemical mechanism of metabolism on reducing blood lipid, by oral administration of egg yolk in rats. A total of 36 Sprague Dawley male rats were randomized into four treatment groups, according to a randomized block design. Each group was further divided into three repeat cages, with each repeat cage comprising of 3 rats. The animals were orally administered with egg yolk once a day, while feeding the same purified pellet diet for 6 weeks. The four treatment groups were: C(control, saline 1.0 g), T1(pork belly oil 1.0 g), T2(egg yolk 1.0 g), T3(pork belly oil 1.0 g and egg yolk 1.0 g alternating every week). The measured parameters in each group are listed as follows in the order of highest to the lowest: daily average gain of body weight(T1>T3>T2>C); blood triglyceride and total cholesterol(T1>C>T3>T2) HDL-C (T2>C>T3>T1); and LDL-C (T1>T3>C>T2). AST and ALT, which are the index of liver function, were the highest in T1 but was lowest in T2. The weights of the liver, spleen, and kidney, except for the abdominal fat, showed no significant difference. The weight of abdominal fat was the highest in T1, but there were no significant difference among C, T2, and T3. The HMG-CoA reductase activity was the highest in T1 followed by T3, C but T2 was lowest. The daily fecal excretions of the total sterol, neutral sterol and acid sterol was highest in T2 but lowest in T1. The results of this study show that the egg consumption reduces the blood lipid through facilitation of fecal excretions of sterols and inhibition of enzyme activity in cholesterol biosynthesis, in the liver of animal and human.

A New Sterol Regulatory Element Binding Protein, SrbB Is Critical for Hypoxia Adaptation and Virulence in the Human Fungal Pathogen Aspergillus fumigatus

  • Chung, Dawoon;Barker, Bridget M.;Carey, Charles C.;Merriman, Brittney;Werner, Ernst R.;Lechner, Beatrix E.;Dhingra, Sourabh;Cheng, Chao;Xu, Wenjie;Blosser, Sara J.;Morohashi, Kengo;Mazurie, Aurelien;Mitchell, Thomas K.;Haas, Hubertus;Mitchell, Aaron P.;Cramer, Robert A.
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.15-15
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    • 2015
  • Aspergillus fumigatus is a major cause of invasive aspergillosis (IA), a significant health issue worldwide with high mortality rates up to 95%. Our lab is interested in how A. fumigatus adapts to low oxygen conditions 'hypoxia', which is one of the important host microenvironments. A. fumigatus SrbA is a basic helix-loop-helix (bHLH) transcriptional regulator and belongs to sterol regulatory element binding protein (SREBP) family members. Loss of SrbA completely blocks growth in hypoxia and results in avirulence in murine models of IA suggesting an essential role of SrbA in hypoxia adaptation and virulence in A. fumigatus. We conducted chromatin immunoprecipitation sequencing (ChIP-seq) with A. fumigatus wild type using a SrbA specific antibody, and 97 genes were revealed as SrbA direct targets. One of the 'SrbA regulons' (AFUB_099590) was a putative bHLH transcriptional regulator whose sequence contained a characteristic tyrosine substitution in the basic portion of the bHLH domain of SREBPs. Therefore, we designated AFUB_099590 SrbB. Further characterization of SrbB demonstrated that SrbB is important for radial growth, biomass production, and biosynthesis of heme intermediates in hypoxia and virulence in A. fumigatus. A series of quantitative real time PCR showed that transcription of several SrbA regulons is coordinately regulated by two SREBPs, SrbA and SrbB in hypoxia. This suggests that SrbA and SrbB have both dependent and independent functions in regulation of genes responsible for hypoxia adaptation in A. fumigatus. Together, our data provide new insights into complicated roles of SREBPs in adaptation of host environments and virulence in pathogenic fungi.

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Comparison of RNA Interference-mediated Gene Silencing and T-DNA Integration Techniques for Gene Function Analysis in Chinese Cabbage (RNA Interference 및 T-DNA Integration 방법에 의한 배추 기능유전자 Silencing 효과 비교)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.30 no.6
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    • pp.734-742
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    • 2012
  • To compare RNA interference-mediated gene silencing technique and T-DNA integration for gene function analysis in Chinese cabbage, BrSAMS-knockout (KO) line and BrSAMS-knockdown (KD) line were used. The KO line had lost the function of a Brassica rapa S-adenosylmethionine synthetase (BrSAMS) gene by T-DNA insertion and the KD line had shown down-regulated BrSAMS genes' expression by dsRNA cleavage. From microarray results of the KO and KD lines, genes linked to SAMS such as sterol, sucrose, homogalacturonan biosynthesis and glutaredoxin-related protein, serine/threonine protein kinase, and gibberellin-responsive protein showed distinct differences in their expression levels. Even though one BrSAMS gene in the KO line was broken by T-DNA insertion, gene expression pattern of that line did not show remarkable differences compared to wild type control. However, the KD line obtained by RNAi technique showed prominent difference in its gene expression. Besides, change of polyamine and ethylene synthesis genes directly associated with BrSAMS was displayed much more in the KD line. In the microarray analysis of the KO line, BrSAMS function could not be clearly defined because of BrSAMS redundancy due to the genome triplication events in Brassicaceae. In conclusion, we supposed that gene knock-down method by RNAi silencing is more effective than knock-out method by T-DNA insertion for gene function analysis of polyploidy crops such as Chinese cabbage.

Effect of Glucosylceramides and Sterols Isolated from Agaricus Blazei Extract on Improvement of Skin Cell (신령버섯에서 분리된 Glucosylceramide 및 Sterol의 피부 세포 개선 효과)

  • Kim, Jeong Eun;Lee, So Young;Chang, Yunhee;Jin, Mu Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.2
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    • pp.105-117
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    • 2020
  • Agaricus blazei Murill (Almond mushroom) has many beneficial effects, such as anti-cancer, immuneenhancement, and anti-obesity. Also, its skin benefits have been reported for antioxidant, anti-inflammatory, and whitening. In order to elucidate these effects, many studies have been conducted. In this study, we reconfirmed the skin efficacy of the extract of the mushrooms mushrooms. The Agaricus blazei extract showed inhibition of melanin synthesis, enhancement of collagen synthesis, and up-regulation of gene expression (hyaluronan syntahase-2, 3 and aquaporin-3) at 100 ㎍/mL. and identified the ingredients from the extract. We further investigated them to find an applicability as cosmetic ingredients. The ingredients were confirmed comparison of their spectroscopic data with literature values. They were identified as being ergosterol (1), 5-dihydroergosterol (2), cerevisterol (3), cerebroside B (4), cerebroside D (5), adenosine (6), and benzoic acid (7). Among these compounds, we evaluated skin efficacy for two cerebrosides and three ergosterol derivatives that have not been reported its efficacy. As a result, 5-dihydroergosterol (2) inhibited melanogenesis in B16F10 and promoted collagen biosynthesis in human dermal fibroblast. In addition, cerevisterol (3), cerebroside B (4), and cerebroside D (5) inhibited NO production in RAW 264.7 cell. In particular, cerebroside D (5) increased the expression of hyaluronan synthase-2 and aquaporin-3 genes in HaCaT. These results suggest that Agaricus blazei extract and isolated compounds can be used as cosmetic ingredients.

Functional Ingredient and Their Some Variance in Amaranth and Quinoa (비름(Amaranth)과 명아주(Quinoa) 재배종의 기능성 물질과 변이)

  • Lee, Jae-Hak;Kim, Ki-Jun;Lee, Jung-Il;Lee, Seung-Tack;Ryu, Su-Noh
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.spc1
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    • pp.145-165
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    • 1996
  • Amaranth(Amaranthus spp. L.) and quinoa (Chenpodium quinoa Willd.) are old crops from South, Central America and Central Asia and their grains have been identified as very promising food crops because of their exceptional nutritive value. Squalene is an important ingredient in skin cosmetics and computer disc lubricants as well as bioactive materials such as inhibition of fungal and mammalian sterol biosynthesis, antitumor, anticancer, and immunomodulation. Amaranth has a component called squalene (2,6,10,15,19,23-hexamethyl-2,6,10,14,22-tetraco-sahexaene) about 1/300 of the seed and $5\~8\%$ of its seed oil. Oil and squalene content in amaranth seed were different for the species investigated. Squalene content in seed oil also increased by $15.5\%$ due to puffing and from 6.96 to $8.01\%$ by refining and bleaching. Saponin concentrations in quinoa seed ranged 0.01 to $5.6\%$. Saponins are located in the outer layers of quinoa grain. These layers include the perianth, pericarp, a seed coat layer, and a cuticle like structure. Oleanane-type triterpenes saponins are of great interest because of their diverse pharmacological properties, for instance, anti-inflammatory, antibiotic, contraceptive, and cholesterol-lowering effects. It is known that quinoa contains a number of structurally diverse saponins including the aglycones, oleanolic acid, hederagenin, and phytolaccagenic acid, which are new potential in gredient for pharmacological properties. It is likely that these saponin levels will be considerably affected by genetic, agronomic and environmental factors as well as by processing. With the current enhanced public interest in health and nutrition amaranth and quinoa will most likely remain in the immediate future within the realm of exotic health foods until such time as agricultural production meets the quantities and qualify required by industrial food manufacturers.

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Alteration of Lipid Metabolism by Ginseng Supplements With Different Levels of Vitamin E in High Cholesterol-Fed Rats

  • Do, Kyung-Min;Park, Yong-Bok;Bok, Song-Hae;Lee, Mi-Kyung;Jeong, Tae-Sook;Choi, Myung-Sook
    • Preventive Nutrition and Food Science
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    • v.6 no.1
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    • pp.66-72
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    • 2001
  • Ginseng may have antioxidant and pharmacologic effects similar to those of vitamin E. The interactive effect of ginseng and vitamin E was studied with respect to cholesterol metabolism and the antioxidant status. A ginseng supplement (0.1%, wt/wt) with comparable levels of vitamin E was provided with a high-cholesterol (1%, wt/wt) diet to rats for 5 weeks. The amount of vitamin E included in the ginseng-free and ginseng diets was either a low (low-E) or a normal (normal-E) level. The ginseng supplements significantly (p<0.05) altered the concentrations of plasma triglycerides in both the low-vitamin E group and normal-vitamin E group compared to the each ginseng-free group. The hepatic triglyceride and cholesterol content were not significantly (p>0.05) different between groups regardless of the vitamin E level in the diet. The hepatic HMG-CoA reductase activity was significantly (p<0.05) lowered by the ginseng supplement in both the low-vitamin E and the normal-vitamin E groups compared to the ginseng-free group. The HMG-CoA reductase activity was also significantly (p<0.05) lowered with in increase of the dietary vitamin E in the ginseng-free group. The excretion of fecal neutral sterol was significantly (p<0.05) lower in the normal-E ginseng group than th low-E ginseng-free group. Neither dietary ginseng nor vitamin E significantly changed the hepatic antioxidant enzymes activity. This data indicates that ginseng supplements lower the concentration of plasma triglyceride and hepatic HMG-CoA reductase activity regardless of eh dietary vitamin E level. This information may contribute to understanding the interactive effect of ginseng and vitamin E on cholesterol biosynthesis in high cholesterol-fed rats.

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Regulation of HMG-CoA Reductase mRNA Stability by 25-hydroxycholesterol

  • Park, Jae-Won;Oh, Seung-Min
    • Preventive Nutrition and Food Science
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    • v.5 no.4
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    • pp.184-188
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    • 2000
  • HMG-CoA reductase is th rate-limiting enzyme of cholesterol biosynthesis. As intracellular levels of cholesterol should be regulated elaborately in response to external stimuli an internal needs, the expression of the HMG-CoA reductase gene is regulated intricately at several different levels from transcription to post-translational modification. In this study, we investigated the regulatory mechanism of HMG-CoA reductase gene expression at the post-transcriptional/pre-translational levels in a baby hamster kidney cell line, C100. when 25-hydroxycholesterol was added to cells cultured in medium containing 5% delipidized fetal bovine serum and 25$\mu$M lovastatin, the levels of HMG-CoA reductase mRNA decreased rapidly, which seemed to be due to the increased degradation of reductase mRNA. These suppressive effects of 25-hydroxycholesterol on MG-CoA reductase mRNA levels were blocked by a translation inhibitor, cycloheximide. Similarly, actinomycin D and 5,6-dichloro-1-$\beta$-D-ribofuranosylbenzimidazole, transcription inhibitors, blocked the 25-hydroxycholesterol-mediated degradation of HMG-CoA reductase mRNA. These results indicate that new protein/RNA synthesis is required for the degradation of HMG-CoA reductase mRNA. In addition, data from the transfection experiments shows that cis-acting determinants, regulating the stability of reductase mRNA, were scattered in the sequence corresponding to 1766-4313 based on the sequence of Syrian hamster HMG-CoA reductase cDNA. Our data suggests that sterol-mediated destabilization of reductase mRNA might be one of the important regulatory mechanism of HMG-CoA reductase gene expression.

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Molecular Cloning and Characterization of the Yew Gene Encoding Squalene Synthase from Taxus cuspidata

  • Huang, Zhuoshi;Jiang, Keji;Pi, Yan;Hou, Rong;Liao, Zhihua;Cao, Ying;Han, Xu;Wang, Qian;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.40 no.5
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    • pp.625-635
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    • 2007
  • The enzyme squalene synthase (EC 2.5.1.21) catalyzes a reductive dimerization of two farnesyl diphosphate (FPP) molecules into squalene, a key precursor for the sterol and triterpene biosynthesis. A full-length cDNA encoding squalene synthase (designated as TcSqS) was isolated from Taxus cuspidata, a kind of important medicinal plants producing potent anti-cancer drug, taxol. The full-length cDNA of TcSqS was 1765 bp and contained a 1230 bp open reading frame (ORF) encoding a polypeptide of 409 amino acids. Bioinformatic analysis revealed that the deduced TcSqS protein had high similarity with other plant squalene synthases and a predicted crystal structure similar to other class I isoprenoid biosynthetic enzymes. Southern blot analysis revealed that there was one copy of TcSqS gene in the genome of T. cuspidata. Semi-quantitative RT-PCR analysis and northern blotting analysis showed that TcSqS expressed constitutively in all tested tissues, with the highest expression in roots. The promoter region of TcSqS was also isolated by genomic walking and analysis showed that several cis-acting elements were present in the promoter region. The results of treatment experiments by different signaling components including methyl-jasmonate, salicylic acid and gibberellin revealed that the TcSqS expression level of treated cells had a prominent diversity to that of control, which was consistent with the prediction results of TcSqS promoter region in the PlantCARE database.