• Title/Summary/Keyword: pathogenesis-related protein

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Development of a Screening System for Plant Defense-Inducing Agent using Transgenic Tobacco Plant with PR-1a Promoter and GUS Gene

  • Oh, Sang-Keun;Lee, Seon-Woo;Kwon, Suk-Yoon;Choi, Do-Il
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.288-292
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    • 2005
  • Pathogenesis-related protein-1a (PR-1a) is strongly induced in tobacco plants by pathogen attack, exogenous salicylic acid (SA) application and by other developmental processes. In order to develop a rapid screening system for the selection of plant defense-inducing compounds originated from various sources, we have transformed tobacco Samsun NN plants with a chimeric construct consisting of GUS $(\beta-glucuronidase)$. In the $T_1$ generation, three transgenic lines having stable GUS expression were selected for further promoter analysis. Using GUS histochemical assay, we observed strong GUS induction driven by PR-1a promoter in PR1a-GUS transgenic tobacco leaves in response to the exogenous application of SA or benzol (1,2,3) thiadiazole-7-carbothioic acid S-methyl ester (BTH), a SA­derivative compound. In addition, GUS expression was maintained locally or systemically in PR1a-GUS transgenic line $\#5\;T_2$ generation) until after 3 days when they were treated with same chemicals. Our results suggested that the PR1a-GUS reporter gene system in tobacco plants may be applicable for the large-scale screening of defense-inducing substances.

Bacterial Multiplications and Electrophoretic Patterns of Soluble Proteins in Compatible and Incompatible Interactions of Pepper Leaves with Xanthomonas campestirs pv. vesicatoria (Xanthomonas campestris pv. vesicatoria에 감염된 고추잎의 친화적, 불친화적 반응에서 세균증식과 수용성 단백질의 전기영동 패턴)

  • 이연경;김영진;황병국
    • Korean Journal Plant Pathology
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    • v.10 no.4
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    • pp.305-313
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    • 1994
  • Typically susceptible lesions were developed on pepper (cv. Hanbyul) leaves inoculated with the compatible strains Ds 1 of Xanthomonas campestris pv. vesicatoria. The lesions appeared first water-soaked and then turned yellow with a chlorotic area. In contrast, the leaves inoculated with the incompatible strain 81-23 initially turned yellow and then developed local necrosis. Multiplication of x. c. pv. vesicatoria in pepper leaves also were distinctly different between the two strains. The strain Ds 1 multiplied more greatly than did the strain 81-23 in the infected leaves. X. c. pv. vesicatoria infection of pepper leaves induced the synthesis of soluble proteins, especially more greatly in the compatible than in the incompatible interactions. Some pathogenesis-related (PR) proteins were detected in the intercellular washing fluid (IWF) and extracts of the infected pepper leaves. In particular, the 32 kDa protein on SDS-PAGE gels appeared intensely in the incompatible interaction. In contrast, some proteins with moluecular masses of 65, 71, and 75 kDa disappeared in the infected pepper leaves. Isoelectric focusing could identify the pIs of soluble proteins in infected pepper leaves. The accumulation of the IWF from infected leaves was more conspicuous in the incompatible than the compatible interaction. These results suggest that some extremely acidic and basic proteins were induced and accumulated in the intercellular spaces of infected pepper leaves.

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Construction of a High-Quality Yeast Two-Hybrid Library and Its Application in Identification of Interacting Proteins with Brn1 in Curvularia lunata

  • Gao, Jin-Xin;Jing, Jing;Yu, Chuan-Jin;Chen, Jie
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.108-114
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    • 2015
  • Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.

Oxidative Modification of Neurofilament-L Induced by Endogenous Neurotoxin, Salsolinol

  • Kang, Jung-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.32 no.9
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    • pp.3421-3424
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    • 2011
  • The endogenous neurotoxin, 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (salsolinol), has been considered a potential causative factor for the pathogenesis of Parkinson's disease (PD). In this study, we examined oxidative modification of neurofilament-L (NF-L) induced by salsolinol. When disassembled NF-L was incubated with salsolinol, the aggregation of protein was increased with the concentration of sasolinol. The formation of carbonyl compound was obtained in salsolinol-mediated NF-L aggregates. This process was protected by free radical scavengers, such as N-acetyl-L-cysteine and glutathione. These results suggest that the aggregation of NF-L is mediated by salsolinol via the generation of free radicals. We also investigated the effects of copper ion on salsolinol-mediated NF-L modification. In the presence of copper ions, salsolinol enhanced the modification of NF-L. We suggest that salsolinol might be related to abnormal aggregation of NF-L which may be involved in the pathogenesis of neurodegenerative diseases and related disorders.

New insight into transglutaminase 2 and link to neurodegenerative diseases

  • Min, Boram;Chung, Kwang Chul
    • BMB Reports
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    • v.51 no.1
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    • pp.5-13
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    • 2018
  • Formation of toxic protein aggregates is a common feature and mainly contributes to the pathogenesis of neurodegenerative diseases (NDDs), which include amyotrophic lateral sclerosis (ALS), Alzheimer's, Parkinson's, Huntington's, and prion diseases. The transglutaminase 2 (TG2) gene encodes a multifunctional enzyme, displaying four types of activity, such as transamidation, GTPase, protein disulfide isomerase, and protein kinase activities. Many studies demonstrated that the calcium-dependent transamidation activity of TG2 affects the formation of insoluble and toxic amyloid aggregates that mainly consisted of NDD-related proteins. So far, many important and NDD-related substrates of TG2 have been identified, including $amlyoid-{\beta}$, tau, ${\alpha}-synuclein$, mutant huntingtin, and ALS-linked trans-activation response (TAR) DNA-binding protein 43. Recently, the formation of toxic inclusions mediated by several TG2 substrates were efficiently inhibited by TG2 inhibitors. Therefore, the development of highly specific TG2 inhibitors would be an important tool in alleviating the progression of TG2-related brain disorders. In this review, the authors discuss recent advances in TG2 biochemistry, several mechanisms of molecular regulation and pleotropic signaling functions, and the presumed role of TG2 in the progression of many NDDs.

Overexpression of a Pathogenesis-Related Protein 10 Enhances Biotic and Abiotic Stress Tolerance in Rice

  • Wu, Jingni;Kim, Sang Gon;Kang, Kyu Young;Kim, Ju-Gon;Park, Sang-Ryeol;Gupta, Ravi;Kim, Yong Hwan;Wang, Yiming;Kim, Sun Tae
    • The Plant Pathology Journal
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    • v.32 no.6
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    • pp.552-562
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    • 2016
  • Pathogenesis-related proteins play multiple roles in plant development and biotic and abiotic stress tolerance. Here, we characterize a rice defense related gene named "jasmonic acid inducible pathogenesis-related class 10" (JIOsPR10) to gain an insight into its functional properties. Semi-quantitative RT-PCR analysis showed up-regulation of JIOsPR10 under salt and drought stress conditions. Constitutive over-expression JIOsPR10 in rice promoted shoot and root development in transgenic plants, however, their productivity was unaltered. Further experiments exhibited that the transgenic plants showed reduced susceptibility to rice blast fungus, and enhanced salt and drought stress tolerance as compared to the wild type. A comparative proteomic profiling of wild type and transgenic plants showed that overexpression of JIOsPR10 led to the differential modulation of several proteins mainly related with oxidative stresses, carbohydrate metabolism, and plant defense. Taken together, our findings suggest that JIOsPR10 plays important roles in biotic and abiotic stresses tolerance probably by activation of stress related proteins.

A study on Histologically Change of the Skin and Liver in Skin Burn (피부화상에 의한 피부 및 간의 조직학적 변화에 관한 연구)

  • Kim, Han-Soo;Kim, Sang-Soo;Kim, Yong-Kwon
    • Journal of Korean Physical Therapy Science
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    • v.8 no.1
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    • pp.923-934
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    • 2001
  • In order to investigate a pathogenesis of liver damage induced by skin burn, thermal injury was induced by scald burn on entirely dorsal surface in rats (total body surface area 30%) except for inhalated injury. At 5 and 24 h after scald bum, biochemical assay and morphological changes in skin tissue, serum and liver tissue were examined. The effects of bum injury on the levels of glutathione, lipid peroxide and on the activities of oxygen free radical generating and scavenging enzymes have been determined in association with observing of histologic and ultrastructural changes, measuring the protein concentration in plasma, and counting the number of intravascular polymorphonuclear leukocytes. The activity of xanthine oxidase, an enzyme of oxygen free radical generating system. was elevated (p<0.01) in serum, but not in skin and in liver tissue. Futhermore, thermal injury decreased not only the protein concentration in plasma but also the number of leukocytes, that indicates induction of edema formation with protein exudation and inflammation by neutrophil infiltration into the internal organs. These data suggest that acute dermal scald burn injury leads to liver damage, that is related to elevation of xanthine oxidase activity in serum. Xanthine oxidase may be a key role in the pathogenesis of liver damage induced by skin burn.

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Beta-amyloid peptide degradation by aminopeptidase and its functional role in Alzheimer's disease pathogenesis

  • AhnJo, Sang-Mee
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2006.04a
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    • pp.77-90
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    • 2006
  • Beta-amyloid peptide (A$\beta$) is a major component of senile plaques and its aggregation is considered to play a critical role in pathogenesis of Alzheimer's disease (AD). Aggregation of A$\beta$ could result from both increased synthesis and decreased degradation of A$\beta$. Our laboratory is interested in understanding the mechanism of A$\beta$ degradation in brain. Recently our laboratory identified a bacterial gene (SKAP) from Streptomyces sp KK565 whose protein product has an activity to cleave A$\beta$ and thus reduce the A$\beta$-induced neurotoxicity. The sequence analysis showed that this gene was closely related to aminopeptidase. Maldi-Tof analysis showed that the recombinant SKAP protein expressed in E. coli cleaves both A$\beta$ 40 and A$\beta$ 42 at the N-terminal of A$\beta$ while an aminopeptidase from Streptomyces griseus (SGAP) cleaves at the C-terminal. We also identified a mammalian homolog of SKAP and the recombinant mammalian protein expressed in Sf-9 insect cells showed a similar proteolytic activity to SGAP, cutting A$\beta$ at the C-terminus. I well discuss the detailed mechanism of the enzyme action and its functional implication in AD.

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C-Reactive Protein a Promising Biomarker of COVID-19 Severity

  • Fazal, Muntaha
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.3
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    • pp.201-207
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    • 2021
  • The 2019 coronavirus outbreak poses a threat to scientific, societal, financial, and health resources. The complex pathogenesis of severe acute respiratory syndrome coronavirus centers on the unpredictable clinical progression of the disease, which may evolve abruptly and result in critical and life-threatening clinical complications. Effective clinical laboratory biomarkers that can classify patients according to risk are essential for ensuring timely treatment, and an analysis of recently published studies found cytokine storm and coagulation disorders were leading factors of severe COVID-19 complications. The following inflammatory, biochemical, and hematology biomarkers markers have been identified in COVID-19 patients; neutrophil to lymphocyte ratio, c-reactive protein, procalcitonin, urea, liver enzymes, lactate dehydrogenase, serum amyloid A, cytokines, d-dimer, fibrinogen, ferritin, troponin, creatinine kinase, and lymphocyte, leukocyte, and platelet counts. These factors are predictors of disease severity and some are involved in the pathogenesis of COVID-19. CRP is an acute-phase, non-specific serological biomarker of inflammation and infection and is related to disease severities and outcomes. In the present study, CRP levels were found to rise dramatically among COVID-19 patients, and our findings suggest CRP could be utilized clinically to predict COVID-19 prognosis and severity even before disease progression and the manifestation of clinical symptoms.