• Title/Summary/Keyword: biochemical pathway

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Role of plastidic glucose transporter in source metabolism of Arabidopsis

  • Lee, Youn-Hyung;Hong, Soon-Won;Lee, Jang-Wook;Bhoo, Seong-Hee;Jeon, Jong-Seong;Hahn, Tae-Ryong
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.9-21
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    • 2005
  • To study the biochemical and physiological role of the plastidic glucose transporter (pGlcT) in carbohydrate metabolism, we characterized transgenic plants with mutations in the pGlcT gene (GT), gt-1 and gt-2, as well double mutants of GT and the maltose transporter (MEX1) and GT and the triose phosphate/phosphate translocator (TPT), GT and the cytosolic fructose-1,6-bisphosphatase gene (cFBP), and MEX1 and TPT, gt-1/mex2, gt-1/tpt-2, gt-1/cfbp-1, mex1-1/tpt-2, respectively. Compared to the wild type, all mutants except the gt-1/cfbp-1 mutant lines displayed higher starch accumulation and higher levels of maltose. Starch accumulation is due to a decrease in starch turnover, leading to an imbalance between the rates of synthesis and degradation. Sucrose levels of gt alleles were higher than those in wild-type plants during the light period, suggesting possible nightly supplementation via the maltose transport pathway to maintain proper carbohydrate partitioning in the plant leaves. The gt plants displayed less growth retardation than mex1-1 mutant and gt-1/mex2 double mutant displayed accumulativesevere growth retardation as compared to individual gt-1 and mex1-1 mutants, implying that the maltose transporter-mediated pathway is a major route for carbohydrate partitioning at night. The gt-1/tpt-2, mex1-1/tpt-2 and gt-1/cfbp-1 double mutants had retarded growth and low chlorophyll content to differing degrees, indicating that photosynthetic capacity had diminished. Interestingly, the gt-1/tpt-2 line displayed a glucose-insensitive phenotype and higher germination rates than wild type, suggesting its involvement not only in carbon partitioning, but also in the sugar signaling network of the pGlcT and TPT.

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Genetic and biochemical evidence for redundant pathways leading to mycosporine-like amino acid biosynthesis in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024

  • Geraldes, Vanessa;de Medeiros, Livia Soman;Lima, Stella T.;Alvarenga, Danillo Oliveira;Gacesa, Ranko;Long, Paul F.;Fiore, Marli Fatima;Pinto, Ernani
    • ALGAE
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    • v.35 no.2
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    • pp.177-187
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    • 2020
  • Cyanobacteria have been widely reported to produce a variety of UV-absorbing mycosporine-like amino acids (MAAs). Herein, we reported production of the unusual MAA, mycosporine-glycine-alanine (MGA) in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024 using a newly developed UHPLC-DAD-MS/HRMS (ultra-high performance liquid chromatography-diode array detection-high resolution tandem mass spectrometry) method. MGA had previously been first identified in a red-algae, but S. torques-reginae strain ITEP-024 is the first cyanobacteria to be reported as an MGA producer. Herein, the chemical structure of MGA is fully elucidated from one-dimensional / two-dimensional nuclear magnetic resonance and HRMS data analyses. MAAs are unusually produced constitutively in S. torques-reginae ITEP-024, and this production was further enhanced following UV-irradiance. It has been proposed that MAA biosynthesis proceeds in cyanobacteria from the pentose phosphate pathway intermediate sedoheptulose 7-phosphate. Annotation of a gene cluster encoded in the genome sequence of S. torques-reginae ITEP-024 supports these gene products could catalyse the biosynthesis of MAAs. However, addition of glyphosate to cultures of S. torques-reginae ITEP-024 abolished constitutive and ultra-violet radiation induced production of MGA, shinorine and porphyra-334. This finding supports involvement of the shikimic acid pathway in the biosynthesis of MAAs by this species.

Proton Magnetic Resonance Spectroscopic Changes of the Primary Motor Cortex and Supplementary Motor Area in Hemiparetic Patients with Corticospinal Tract Injury due to Deep Intracerebral Hematoma

  • Yang, Dong-Joon;Son, Byung-Chul;Baik, Hyun-Man;Lee, Sang-Won;Sung, Jae-Hoon;Choe, Bo-Young
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2005.04a
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    • pp.32-34
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    • 2005
  • Purpose: To investigate the metabolic changes in the motor and motor association cortices following axonal injury in the internal capsule that was caused by deep intracerebral hematoma. Materials and Methods: Using proton magnetic resonance spectroscopy (1H MRS), the authors studied the primary motor cortices (M-1) and supplementary motor areas (SMA) of 9 hemiparetic patients with documentable hemiparesis of varying severity, and we studied 10 normal volunteers as controls. To measure the M-1 and SMA biochemical changes, 4 separate single volumes of interest (VOIs) were located bilaterally in the affected and unaffected hemisphere (AH and UH). Results: 1H MRS provided a neuronal and axonal viability index by measuring levels of N-acetylaspartate (NAA) and creatine/phosphocreatine (Cr). The M-1/SMA NAA/Cr ratios of the AH and UH in patients, and the AH and normal volunteers were compared. The NAA/Cr ratios of the M-1 and SMA in AH, and the SMA in UH were significantly lower than those of normal volunteers. Conclusion: These 1H MRS findings indicate that axonal injury in the descending motor pathway at the level of internal capsule could induce metabolic changes in the higher centers of the motor pathway.

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"A La Recherche" of Functions for the Spore Protein SASP-E from Bacillus subtilis

  • Ruzal, Sandra M.;Bustos, Patricia L.;Sanchez-Rivas, Carmen
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.15-21
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    • 2013
  • We previously observed that Bacillus subtilis spores from sspE mutants presented a lower germination capacity in media containing high salt concentrations (0.9M NaCl). This deficiency was attributed to the absence of SASP-E (gamma-type small-acid-soluble protein), rich in osmocompatible amino acids released by degradation. Herein we observed that, in addition, this mutant spore presented a reduced capacity to use L-alanine as germinant (L-ala pathway), required longer times to germinate in calcium dipicolinate ($Ca^{2+}$-DPA), but germinated well in asparagine, glucose, fructose, and potassium chloride (AGFK pathway). Moreover, mild sonic treatment of mutant spores partially recovered their germination capacity in L-ala. Spore qualities were also altered, since sporulating colonies from the sspE mutant showed a pale brownish color, a higher adherence to agar plates, and lower autofluorescence, properties related to their spore coat content. Furthermore, biochemical analysis showed a reduced partition in hexadecane and a higher content of $Ca^{2+}$-DPA when compared with its isogenic wild-type control. Coat protein preparations showed a different electrophoretic pattern, in particular when detected with antibodies against CotG and CotE. The complementation with a wild-type sspE gene in a plasmid allowed for recovering the wild-type coat phenotype. This is the first report of a direct involvement of SASP-E in the spore coat assembly during the differentiation program of sporulation.

Dietary effects of sophorolipids on nutrient bioavailability and intestinal microenvironments in broiler chickens

  • Min-Jin, Kwak;Min Young, Park;Ki-Peum, Sung;Hanbae, Lee;Kwang-Youn, Whang;Younghoon, Kim
    • Journal of Animal Science and Technology
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    • v.64 no.6
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    • pp.1092-1104
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    • 2022
  • Using antibiotics as growth promoter has been banned in poultry feed industry, thus various researchers try to seek an alternative to replace the growth-promoting antibiotics. In this study, we aimed to evaluate the growth performance via intestinal nutrient utilization and cecal microbial composition of broiler after dietary supplementation with most commonly using antibiotics, zinc bacitracin, and sophorolipid. A total of 180 1-day-old chicks were randomly assigned, and dietary treatment was as follow: CON, basal diet; ZB, 100 ppm of zinc bacitracin supplemented diet; and SPL, 250 ppm of sophorolipid supplemented diet. Their growth performance was evaluated and the samples of blood, small intestine, and ileal and cecal digesta were collected for biochemical, histological, and genomic analyses. The body weight and average daily gain of 7-day-old chicks were higher in ZB and those in overall experimental period were improved by ZB and SPL supplementation (p < 0.05). Their intestinal characteristics were not affected by dietary treatments in duodenum and ileum. Nonetheless, villus height was increased by SPL supplementation in jejunum (p < 0.05). Moreover, dietary SPL supplementation could down-regulate the expression level of pro-inflammatory cytokine, IL-1β (p < 0.05). mRNA levels of lipid and protein transporters did not differ among the treatments, however, relative expression levels of carbohydrate transporters, GLUT2 and SGLT1 were increased in broiler chicken's jejumum fed zinc bacitracin and sophorolipid supplemented diets (p < 0.05). Dietary zinc bacitracin supplementation could increase the population of Firmicutes in phylum level, and the portion of Turiciacter in genus level. On the other hands, the portion of Faecalibacterium was increased by dietary SPL supplementation compared to the other treatments. Our findings suggest that SPL supplementation improves growth performance through enhanced carbohydrate utilization capacity via improvement of gut morphological status and modulation of the cecal microbial population of broilers.

Microtubule Acetylation-Specific Inhibitors Induce Cell Death and Mitotic Arrest via JNK/AP-1 Activation in Triple-Negative Breast Cancer Cells

  • Suyeon Ahn;Ahreum Kwon;Youngsoo Oh;Sangmyung Rhee;Woo Keun Song
    • Molecules and Cells
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    • v.46 no.6
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    • pp.387-398
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    • 2023
  • Microtubule acetylation has been proposed as a marker of highly heterogeneous and aggressive triple-negative breast cancer (TNBC). The novel microtubule acetylation inhibitors GM-90257 and GM-90631 (GM compounds) cause TNBC cancer cell death but the underlying mechanisms are currently unknown. In this study, we demonstrated that GM compounds function as anti-TNBC agents through activation of the JNK/AP-1 pathway. RNA-seq and biochemical analyses of GM compound-treated cells revealed that c-Jun N-terminal kinase (JNK) and members of its downstream signaling pathway are potential targets for GM compounds. Mechanistically, JNK activation by GM compounds induced an increase in c-Jun phosphorylation and c-Fos protein levels, thereby activating the activator protein-1 (AP-1) transcription factor. Notably, direct suppression of JNK with a pharmacological inhibitor alleviated Bcl2 reduction and cell death caused by GM compounds. TNBC cell death and mitotic arrest were induced by GM compounds through AP-1 activation in vitro. These results were reproduced in vivo, validating the significance of microtubule acetylation/JNK/AP-1 axis activation in the anti-cancer activity of GM compounds. Moreover, GM compounds significantly attenuated tumor growth, metastasis, and cancer-related death in mice, demonstrating strong potential as therapeutic agents for TNBC.

Physiological and Biochemical Analyses of Rice Sensitivities to UVB Radiation

  • Hidema, Jun;Kumagai, Tadashi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.162-165
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    • 2002
  • Rice is widely cultivated in various regions throughout Asia. Over a five-year period, we investigated the effects of supplemental UVB radiation on the growth and yield of Japanese rice cultivars in the field. The findings of that study indicated that supplemental UVB radiation has inhibitory effects on the growth and grain development. Furthermore, we investigated the sensitivity to UVB radiation of rice cultivars of 5 Asian rice ecotypes, and found that rice cultivars vary widely in UVB sensitivity. The aim of our study is improving UVB resistance in plants by bioengineering or breeding programs. In order to make it, there is need to find the molecular origin of the sensitivity to UVB. Cyclobutane pyrimidine dimer (CPD) is major UV-induced DNA lesions. Plants possess two mechanisms to cope with such DNA damage. The first is the accumulation of UV-absorbing compounds. Our previous data showed that the steady-state CPD levels in leaves of rice grown under chronic radiation in any culture were not so greatly influenced by the increased UV-absorbing compounds content, although there was a significant positive correlation between the CPD levels induced by challenge UVB exposure and the UV-absorbing compounds content. The other is the repair of DNA damage. Photorepair is the major pathway in plants for repairing CPD. We found that the sensitivity to UVB could seriously correlate with the low ability in CPD photorepair in rice plants. These results suggest that photo lyase might be an excellent candidate for restoration by way of selective breeding or engineering in rice.

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Isolation of soybean mutants with high and low inorganic phosphorus

  • Sundaramoorthy, Jagadeesh;Seo, Yean Joo;Park, Gyu Tae;Lee, Jeong-Dong;Park, Soon-Ki;Seo, Hak Soo;Song, Jong Tae
    • Journal of Applied Biological Chemistry
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    • v.59 no.3
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    • pp.261-264
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    • 2016
  • In soybean (Glycine max (L.) Merr.) seeds, phosphorus (P) is primarily stored in the form of phytate, which is generally indigestible by monogastric animals such as human, pig, poultry, and fish. Thus, this study was conducted to isolate soybean mutants with high available P. Inorganic P content was assessed in a total of 1,266 ethyl methanesulfonate (EMS) $M_4$ lines from the Pungsannamul cultivar. Among the tested lines, four EMS lines (PE379, PE432, PE2205, and PE2503) showed higher mean inorganic P ($1.21-1.56gkg^{-1}$) than did the Pungsannamul cultivar ($0.90gkg^{-1}$). Additionally, six EMS lines (PE718, PE828, PE1466, PE1552, PE3378, and PE3386) showed lower mean inorganic P ($0.38-0.60gkg^{-1}$). The high inorganic P mutants isolated in this study will be further investigated for phytate and total P levels. Moreover, the high and low inorganic P lines will be utilized in a future study of the biochemical pathway of phytate.

Semantic Modeling for SNPs Associated with Ethnic Disparities in HapMap Samples

  • Kim, HyoYoung;Yoo, Won Gi;Park, Junhyung;Kim, Heebal;Kang, Byeong-Chul
    • Genomics & Informatics
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    • v.12 no.1
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    • pp.35-41
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    • 2014
  • Single-nucleotide polymorphisms (SNPs) have been emerging out of the efforts to research human diseases and ethnic disparities. A semantic network is needed for in-depth understanding of the impacts of SNPs, because phenotypes are modulated by complex networks, including biochemical and physiological pathways. We identified ethnicity-specific SNPs by eliminating overlapped SNPs from HapMap samples, and the ethnicity-specific SNPs were mapped to the UCSC RefGene lists. Ethnicity-specific genes were identified as follows: 22 genes in the USA (CEU) individuals, 25 genes in the Japanese (JPT) individuals, and 332 genes in the African (YRI) individuals. To analyze the biologically functional implications for ethnicity-specific SNPs, we focused on constructing a semantic network model. Entities for the network represented by "Gene," "Pathway," "Disease," "Chemical," "Drug," "ClinicalTrials," "SNP," and relationships between entity-entity were obtained through curation. Our semantic modeling for ethnicity-specific SNPs showed interesting results in the three categories, including three diseases ("AIDS-associated nephropathy," "Hypertension," and "Pelvic infection"), one drug ("Methylphenidate"), and five pathways ("Hemostasis," "Systemic lupus erythematosus," "Prostate cancer," "Hepatitis C virus," and "Rheumatoid arthritis"). We found ethnicity-specific genes using the semantic modeling, and the majority of our findings was consistent with the previous studies - that an understanding of genetic variability explained ethnicity-specific disparities.

Site-Directed Mutagenesis of Two Cysteines (155, 202) in Catechol 1,2-dioxygenase $I_1$ of Acinetobacter lwoffii K24

  • Kim, Seung-Il;Kim, Soo-Jung;Leem, Sun-Hee;Oh, Kye-Heon;Kim, Soo-Hyun;Park, Young-Mok
    • BMB Reports
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    • v.34 no.2
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    • pp.172-175
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    • 2001
  • Catechol 1,2-dioxygenase $I_1$ ($CDI_1$) is the first enzyme of the $\beta$-ketoadipate pathway in Acinetobacter lowffii K24. $CDI_1$ has two cysteines (155, 202) and its enzyme activity is inhibited by the cysteine inhibitor, $AgNO_3$. Two mutants, $CDI_1$ C155V and $CDI_1$ C202V, were obtained by site-directed mutagenesis. The two mutants were overexpressed and the mutated amino acid residues (Cys$\rightarrow$Val) were characterized by peptide mapping and amino acid sequencing. Interestingly, $CDI_1$ C155V was inhibited by $AgNO_3$, whereas $CDI_1$ C202V was not inhibited. This suggests that $Cys^{202}$ is the sole inhibition site by $AgNO_3$ and is close to the active site of the enzyme. However, the results of the biochemical assay of mutated $CDI_1s$ suggest that the two cysteines are not directly involved in the activity of the catechol 1,2-dioxygenase of $CDI_1$.

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