• Title/Summary/Keyword: functional family

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Genetic Screening for Plant Cell Death Suppressors and Their Functional Analysis in Plants

  • Yun, Dae-Jin
    • Proceedings of the Korean Society of Life Science Conference
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    • 2005.04a
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    • pp.23-36
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    • 2005
  • Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed In yeast. To investigate whether .Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various orgarusms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs In detail. PBIl is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Bax-induced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower lovels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. H$_{2O2}$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of H2O2 treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased In the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 i'n vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation In situ. Thus, AtNDPK2 appears to play a novel regulatory role in H2O2-mediated MAPK signaling in plants.

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Effect of Starter Cultures on the Antioxidant Activities of Allium hookeri Root-Hot Water Extract (발효 균주에 따른 삼채뿌리 열수추출물의 항산화능 비교연구)

  • Jang, Ja-Young;Lee, Jieun;Choi, Eun-Ji;Choi, Hak-Jong;Oh, Young Jun;Lee, Sung Hyun;Kim, Hyun Ju
    • Korean journal of food and cookery science
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    • v.31 no.1
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    • pp.98-102
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    • 2015
  • Allium hookeri, a member of the onion family, has long been mainly cultivated for food and medicinal use in Southeast Asian countries, owing to its various biological properties. However, no studies of the anti-oxidative effects of fermented A. hookeri root extracts have been conducted to date. Therefore, this study investigated the effect of different starter cultures on the antioxidant activities of hot water extract of A. hookeri root by using the following five strains: Lactobacillus acidophilus, Bifidobacterum longum, Enterococcus faecium, Streptococcus thermophilus, and Saccharomyces cerevisiae. DPPH and ABTS radical scavenging activity, total phenolic acid contents, and total antioxidant capacity were higher in the hot water extract of A. hookeri root fermented with starter cultures than those of A. hookeri root. Among hot water extract of A. hookeri root fermented with starter cultures, fementing with S. cerevisiae showed the highest antioxidant activities. The results of this study provide new evidence of the anti-oxidative properties of A. hookeri root with starter cultures, indicating that it may be highly valuable as a natural product owing to its high-quality functional components.

Role of Oxidative Stress and Mitochondria in Parkinson's Disease

  • Jin, Son-Hyeung
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2007.04a
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    • pp.147-153
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    • 2007
  • Central to developing new treatment strategies for late onset sporadic Parkinson's disease (PD) and early onset familial PD is resolving the enigma of the specific vulnerability exhibited by substantia nigra dopamine (DA) neurons despite multiple risk factors. Neuropathological evidence from both human and experimental models of PD firmly supports a significant role for oxidative stress (OS) and mitochondrial dysfunction in the death of nigral DA neurons. Largely unknown are the genes underlying selective susceptibility of nigral DA neuron to OS and mitochondrial dysfunction and how they effect nigral DA cell death. To overcome the paucity of nigral DA neurons as well as the dilution effect of non-DA cells in brain tissues, we have developed wild type DA cell line model, SN4741 and mutant DJ-1 (-/-) DA cells, appropriate for microarray analysis and differential mitochondrial proteomics. Mutations in the DJ-1 gene (PARK7), localized in cytoplasm and mitochondria, cause autosomal recessive early onset PD. Through microarray analysis using SN4741 cells followed by validation tests, we have identified a novel phylogenically conserved neuroprotective gene, Oxi-a, which is specifically expressed in DA neurons. The knockdown of the gene dramatically increased vulnerability to as. Importantly as down-regulated the expression level of the gene and recovery of its expression via transient transfection exerted significant neuroprotection against as insult. We also have identified altered expression of mitochondrial proteins and other familial PD genes in DJ-1 (-/-) mutant cells by differential mitochondrial proteomics. In DJ-1 (-/-) cells the knockdown of the other familial PD genes (Parkin and PINK1) dramatically increased susceptibility to as. Thus, further functional characterization of the Oxi-$\alpha$ gene family and the mitochondrial alteration in the DJ-1 (-/-) cell model will provide the rationale for the neuroprotective therapy against both sporadic and familial PD.

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Isolation and functional characterization of BrUGT gene encoding a UDP-glycosyltransferase from Chinese cabbage (Brassica rapa)

  • Jung, Yu-Jin;Lee, Hye-Jung;Choi, Jang-Sun;Cho, Yong-Gu;Nou, Ill-Sup;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.212-218
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    • 2012
  • Glycosyltransferases are enzymes (EC 2.4) that catalyze the transfer of monosaccharide moieties from activated nucleotide sugar to a glycosyl acceptor molecule which can be a carbohydrate, glycoside, oligosaccharide, or a polysaccharide. In this study, a UDP-glucosyltransferase cDNA was isolated from Brassica rapa using a rapid amplification of cDNA ends (RACE) and subsequently named BrUGT. It has a full-length cDNA of 1,236 bp with 119 bp 5'-untranslated region (UTR), a complete ORF of 834 bp encoding a polypeptide of 277 amino acids (31.19 kDa) and a 3'-UTR of 283 bp. BLASTX analysis hits a catalytic domain of Glycos_transf_1 super family (cl12012) that belongs to the Glycosyltransferases group 1 with tetratricopeptide (TPR) regions located between 165 to 350 bp. Expression analysis showed high mRNA transcripts in pistil, followed by petal, seed and calyx of flower. Moreover, expression analysis of BrUGT in Chinese cabbage seedlings under stresses of cold, salt, PEG, $H_2O_2$, drought and ABA showed elevated mRNA transcript. Furthermore, when BrUGT gene was transformed into rice using pUbi-1 promoter, overexpression was evident among the $T_1$ plants. This study provides insights into the function of BrUGT in plants.

The ABA Effect on the Accumulation of an Invertase Inhibitor Transcript that Is Driven by the CAMV35S Promoter in ARABIDOPSIS

  • Koh, Eun-Ji;Lee, Sung June;Hong, Suk-Whan;Lee, Hoi Seon;Lee, Hojoung
    • Molecules and Cells
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    • v.26 no.3
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    • pp.236-242
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    • 2008
  • Invertase (${\beta}$-D-fructofuranosidase; EC 3.2.1.26) catalyzes the conversion of sucrose into glucose and fructose and is involved in an array of important processes, including phloem unloading, carbon partitioning, the response to pathogens, and the control of cell differentiation and development. Its importance may have caused the invertases to evolve into a multigene family whose members are regulated by a variety of different mechanisms, such as pH, sucrose levels, and inhibitor proteins. Although putative invertase inhibitors in the Arabidopsis genome are easy to locate, few studies have been conducted to elucidate their individual functions in vivo in plant growth and development because of their high redundancy. In this study we assessed the functional role of the putative invertase inhibitors in Arabidopsis by generating transgenic plants harboring a putative invertase inhibitor gene under the control of the CaMV35S promoter. A transgenic plant that expressed high levels of the putative invertase inhibitor transcript when grown under normal conditions was chosen for the current study. To our surprise, the stability of the invertase inhibitor transcripts was shown to be down-regulated by the phytohormone ABA (abscisic acid). It is well established that ABA enhances invertase activity in vivo but the underlying mechanisms are still poorly understood. Our results thus suggest that one way ABA regulates invertase activity is by down-regulating its inhibitor.

Impact of the oral health-related quality of life(OHIP-14) of industrial workers in some area (일부지역 사업장 근로자의 구강건강관련 삶의 질에 미치는 영향)

  • Cheon, Hye-Won
    • Journal of Korean society of Dental Hygiene
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    • v.14 no.6
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    • pp.859-869
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    • 2014
  • Objectives: The purpose of this study was to examine the subjective oral health awareness, oral health knowledge, oral health behavior and OHIP-14 in industrial workers. Methods: A self-reported questionnaire was filled out by 243 workers in Jeonbuk May 7 to June 10, 2014. Except incomplete answers, 230 data were analyzed. The questionnaire consisted of general characteristics of the subjects(sex, age, career, marital status, abd family), oral health recognition characteristics(oral health attention, subjective oral health status, and oral health concern), oral health knowledge, oral health behavior, and oral health related quality of life. The instrument was 14 questions od OHIP-14 including functional restriction(2 questions), physical pain(2 questions), psychological discomfort(2 questions), physical ability decease(2 questions), psychological function decease(2 questions), social activity decrease (2 questions), and social discomfort(2 questions). Cronbach's alpha was 0.949 in this study and it was reliable. Results: Oral health interests showed that 57.8% of the workers had concern for oral health interests and 50.4% perceived that their subjective oral health was moderate status. 55.6% of the workers answered that their oral health status was very worried. Women had more knowledge about the oral health. Those who were in fifties tended to have more knowledge of oral health than the other age groups. Those who had more concern for oral health included female workers, married workers, and workers above 21 years. The concern for oral health made the workers keep good oral health. Higher score of OHIP-14 means good oral health. Conclusions: Good oral health-related quality of life is proportional to continuous management of oral health and subjective oral health status. It is necessary to develop the tailored oral health education program for the workers.

Screening for DPPH Free Radical Scavenging Activities of Autogenous Seaweeds in Jeju Island Using a Electron Spin Resonance (ESR) Spectroscopy (Electron Spin Resonance을 이용한 제주 자생 해조류의 DPPH Free Radical 소거활성 검색)

  • Cha, Seon-Heui;Heo, Soo-Jin;Jeon, You-Jin
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.4
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    • pp.252-259
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    • 2006
  • Extracts which were prepared by four different extractions - 80% methanol extracts (ME) at high ($70^{\circ}C$) and a room temperature ($20^{\circ}C$), respectively and aqueous extracts (AE) at both temperatures with the residue after the methanol extracts - of 10 green, 19 brown and 25 red seaweeds collected in Jeju Island coast were examined for their DPPH free radical scavenging activity using a ESR (electron spin resonance) spectroscopy. A variety of the extracts showed positive scavenging effect against DPPH free radical (except the green seaweeds). Among the extracts, the brown seaweed extracts exhibited the highest scavenging activity. Especially, Sargassum spp. of the brown seaweeds have remarkable scavenging activities - both methanolic and aqueous at the both temperatures ($20^{\circ}C$ and $70^{\circ}C$). On the other hand, ME showed better scavenging activity than AE in the red seaweed extracts. These results indicate that autogenous seaweeds in Jeju will be potential natural antioxidants for functional food compounds.

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Antimicrobial activities and skin barrier improvement effect of Eruca sativa extract (루꼴라(Eruca sativa) 추출물의 항균활성과 피부장벽 개선 효과)

  • Kim, Bora;Kim, Hyun-Soo
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.320-324
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    • 2017
  • Eruca sativa is a rocket plant and a member of the Brassicaceae, which is considered to be an important chemo-preventive plant family. Although Eruca sativa has positive biological effects, the effect of Eruca sativa extract (ES) on improvement of skin barrier function has not been reported. In this study, we investigated the applicability of functional materials by examining a variety of physiological activities of Eruca sativa extract. ES showed anti-microbial activities against Bacillus subtilis, Escherichia coli, and Candida albicans. In particular, antimicrobial activities of ES against B. subtilis was the highest. Additionally, immunohistochemical analysis of protein marker related to keratinocyte differentiation was determined. The treatment by ES (50 mg/L) showed a significant increase of involucrin expression compared with treatment by 0.1% DMSO as a control in skin equivalents, the ES-treated group showed similar level in the expression of involucrin compared to the group treated with the same concentration of WY14643 in $EpiDerm^{TM}$, a three-dimensional model of skin equivalents. These results indicate that ES promotes the expression of protein related to barrier properties of the skin. Therefore, ES may be an effective ingredient for skin barrier improvement.

Crystal structure of CodW in Bacillus Subtilis - the first N-terminal serine pretense

  • Park, Seong-Hwan;Park, Hyun-Ho;Lim, Young-Jun;Kang, Min-Suk;Lim, Byung-Kook;Seong, Ihn-Sik;Jimin Wang;Chung, Chin-Ha;Eom, Soo-Hyun
    • Proceedings of the Korea Crystallographic Association Conference
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    • 2002.11a
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    • pp.25-25
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    • 2002
  • CodWX, encoded by the cod operon in Bacillus subtilis, is a member of the ATP-dependent protease complex family, and is homologous to the eukaryotic 26S proteasome. It consists of two multimeric complexes: two hexameric ATPase caps of CodX and a protease chamber consisting of CodW dodecamer. Prior structural studies have shown that the N-terminal threonine residue is solely functional as a proteolytic nucleophile in ATP-dependent proteases such as HslV and certain β-type subunits of 20S proteasome, which have a primary sequence similarity of -50% and -20% with CodW respectively. Here we present a 3.0 Å resolution crystal structure of CodW, which is the first N-terminal serine protease among the known proteolytic enzymes. In spite of the same fold and the conserved contacts between subunits with HslV in E. coli and H. influenza, this structure shows the five additional residues extending from conserved Thr1 among the other ATP-dependent pretense and extraordinary basic proteolytic chamber.

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Isolation of a Promoter Element that is Functional in Bacillus subtilis for Heterologous Gene Expression

  • Maeng, Chang-Jae;Kim, Hyung-Kwoun;Park, Sun-Yang;Koo, Bon-Tag;Oh, Tae-Kwang;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.85-91
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    • 2001
  • To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

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