• Title/Summary/Keyword: Calmodulin

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Cell Biological Studies on Growth and Development Effect of$Ca^{2+}$ and polyamine of $\beta-glucan$synthetase activity in carrot root protoplast (생체생장에 관한 세포생물학적 연구 당근 뿌리의 원형질체에서 polyamine과 $Ca^{2+}$$\beta-glucan$ synthetase 활성도에 미치는 영향)

  • 이순희
    • Journal of Plant Biology
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    • v.30 no.3
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    • pp.173-179
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    • 1987
  • The effect of polyamine, Ca2+ and calmodulin on GS ($\beta$-glucan synthetase) activity was studied in Daucus carota root. The Ca2+ is shown to have no effect on the GS activity whereas the GS II activity is increased in response to increase in concentration of the Ca2+. When the protoplasts are cultured, for 4 days, the GS II activity increases as a tunction of time and reachs a maximum after 3 days at a time when the network of cellulose microfibrils is known to be synthesized. The effect of the Ca2+ and 1mM spermine on the GS II activity turns out to be synergistic, especially more synergistic at lower concentration of the Ca2+. The GS II activity seems to be enhanced by the Ca2+. The GS II activity in the protoplast treated by the calcium channel blocker, verapamil, turns out to be lower than that of the control. Cumulative results suggest that the Ca2+ stimulates the cell wall regeneration via enhancement of the GS II activity responsible for synthesizing the cell wall component throught synergistic effect with spermine.

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Expression and phosphorylation analysis of soluble proteins and membrane-localised receptor-like kinases from Arabidopsis thaliana in Escherichia coli

  • Oh, Eun-Seok;Eva, Foyjunnaher;Kim, Sang-Yun;Oh, Man-Ho
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.315-321
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    • 2018
  • Molecular and functional characterization of proteins and their levels is of great interest in understanding the mechanism of diverse cellular processes. In this study, we report on the convenient Escherichia coli-based protein expression system that allows recombinant of soluble proteins expression and cytosolic domain of membrane-localised kinases, followed by the detection of autophosphorylation activity in protein kinases. This approach is applied to regulatory proteins of Arabidopsis thaliana, including 14-3-3, calmodulin, calcium-dependent protein kinase, TERMINAL FLOWER 1(TFL1), FLOWERING LOCUS T (FT), receptor-like cytoplasmic kinase and cytoplasmic domain of leucine-rich repeat-receptor like kinase proteins. Our Western blot analysis which uses phospho-specific antibodies showed that five putative LRR-RLKs and two putative RLCKs have autophosphorylation activity in vitro on threonine and/or tyrosine residue(s), suggesting their potential role in signal transduction pathways. Our findings were also discussed in the broader context of recombinant expression and biochemical analysis of soluble and membrane-localised receptor kinases in microbial systems.

Aspergillus caninus (Syn: Phialosimplex caninus): a New Isolate from Field Soils in Korea

  • Adhikari, Mahesh;Gurung, Sun Kumar;Kim, Sang Woo;Lee, Hyun Goo;Ju, Han Jun;Gwon, Byeong Heon;Kosol, San;Bazie, Setu;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.383-392
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    • 2018
  • During the study of indigenous fungal communities in soil samples collected from various field soils in Sancheong, Gyeongsangnam-do, Korea in 2017, several species of Aspergillus were discovered. Aspergillus caninus (KNU17-7) was isolated, identified, and described based on the results from macro and micro morphological characteristics and molecular characterization. Morphologically, it was identified using five different growth media: potato dextrose agar, oatmeal agar, yeast extract sucrose agar, czapek yeast extract agar, and malt extract agar. For the molecular identification, sequencing of internal transcribed spacer, ${\beta}-tubulin$, and calmodulin genes was performed. Based on this characterization, our study isolate was identified as Aspergillus caninus. This fungal species has not been officially reported in Korea before, and we report here with its morphological and molecular phylogenetic characterization.

Fungal Diversity and Enzyme Activity Associated with the Macroalgae, Agarum clathratum

  • Lee, Seobihn;Park, Myung Soo;Lee, Hanbyul;Kim, Jae-Jin;Eimes, John A.;Lim, Young Woon
    • Mycobiology
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    • v.47 no.1
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    • pp.50-58
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    • 2019
  • Agarum clathratum, a brown macroalgae species, has recently become a serious environmental problem on the coasts of Korea. In an effort to solve this problem, fungal diversity associated with decaying A. clathratum was investigated and related ${\beta}$-glucosidase and endoglucanase activities were described. A total of 233 fungal strains were isolated from A. clathratum at 15 sites and identified 89 species based on morphology and a multigene analysis using the internal transcribed spacer region (ITS) and protein-coding genes including actin (act), ${\beta}$-tubulin (benA), calmodulin (CaM), and translation elongation factor (tef1). Acremonium, Corollospora, and Penicillium were the dominant genera, and Acremonium fuci and Corollospora gracilis were the dominant species. Fifty-one species exhibited cellulase activity, with A. fuci, Alfaria terrestris, Hypoxylon perforatum, P. madriti, and Pleosporales sp. Five showing the highest enzyme activities. Further enzyme quantification confirmed that these species had higher cellulase activity than P. crysogenum, a fungal species described in previous studies. This study lays the groundwork for bioremediation using fungi to remove decaying seaweed from populated areas and provides important background for potential industrial applications of environmentally friendly processes.

Penicillium from Rhizosphere Soil in Terrestrial and Coastal Environments in South Korea

  • Park, Myung Soo;Lee, Jun Won;Kim, Sung Hyun;Park, Ji-Hyun;You, Young-Hyun;Lim, Young Woon
    • Mycobiology
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    • v.48 no.6
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    • pp.431-442
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    • 2020
  • Penicillium, the most common genus plays an important ecological role in various terrestrial and marine environments. However, only a few species have been reported from rhizosphere soil. As part of a project to excavate Korean indigenous fungi, we investigated rhizosphere soil of six plants in the forest (terrestrial habitat) and sand dunes (coastal habitat) and focused on discovering Penicillium species. A total of 64 strains were isolated and identified as 26 Penicillium species in nine sections based on morphological characteristics and the sequence analysis of β-tubulin and calmodulin. Although this is a small-scale study in a limited rhizosphere soil, eight unrecorded species and four potential new species have been identified. In addition, most Penicillium species from rhizosphere soil were unique to each plant. Penicillium halotolerans, P. scabrosum, P. samsonianum, P. jejuense, and P. janczewskii were commonly isolated from rhizosphere soil. Eight Penicillium species, P. aurantioviolaceum, P. bissettii, P. cairnsense, P. halotolerans, P. kananaskense, P. ortum, P. radiatolobatum, and P. verhagenii were recorded for the first time in Korea. Here, we provide the detailed morphological description of these unrecorded species.

New Species and Eight Undescribed Species Belonging to the Families Aspergillaceae and Trichocomaceae in Korea

  • Nguyen, Thuong T.T.;Noh, Kyo Jang Kwan;Lee, Hyang Burm
    • Mycobiology
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    • v.49 no.6
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    • pp.534-550
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    • 2021
  • During a survey of fungal diversity associated with insects, mud, soil, and freshwater niches in different areas in Korea, nine interesting fungal strains were isolated. Based on their morphological characteristics and molecular phylogeny analyses, using a combined data set of b-tubulin (BenA), calmodulin (CaM), and second largest subunit of RNA polymerase (RPB2) sequences, the strains CNUFC AM-44, CNUFC JCW3-4, CNUFC S708, CNUFC WT202, CNUFC AS1-29, CNUFC JCW3-5, CNUFC JDP37, and CNUFC JDP62 were identified as Aspergillus alabamensis, A. floridensis, A. subversicolor, Penicillium flavigenum, P. laevigatum, P. lenticrescens, Talaromyces adpressus, and T. beijingensis, respectively. The strain CNUFC JT1301 belongs to series Westlingiorum in section Citrina and is phylogenetically related to P. manginii. However, slow growth when cultivated on CYA, MEA, CREA is observed and the property can be used to easily distinguish the new species from these species. Additionally, P. manginii is known to produce sclerotia, while CNUFC JT1301 strain does not. Herein, the new fungal species is proposed as P. aquadulcis sp. nov. Eight species, A. alabamensis, A. floridensis, A. subversicolor, P. flavigenum, P. laevigatum, P. lenticrescens, T. adpressus, and T. beijingensis, have not been previously reported in Korea. The present study expands the known distribution of fungal species belonging to the families Aspergillaceae and Trichocomaceae in Korea.

Dikkopf-1 promotes matrix mineralization of osteoblasts by regulating Ca+-CAMK2A- CREB1 pathway

  • Hyosun, Park;Sungsin, Jo;Mi-Ae, Jang;Sung Hoon, Choi;Tae-Hwan, Kim
    • BMB Reports
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    • v.55 no.12
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    • pp.627-632
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    • 2022
  • Dickkopf-1 (DKK1) is a secreted protein that acts as an antagonist of the canonical WNT/β-catenin pathway, which regulates osteoblast differentiation. However, the role of DKK1 on osteoblast differentiation has not yet been fully clarified. Here, we investigate the functional role of DKK1 on osteoblast differentiation. Primary osteoprogenitor cells were isolated from human spinal bone tissues. To examine the role of DKK1 in osteoblast differentiation, we manipulated the expression of DKK1, and the cells were differentiated into mature osteoblasts. DKK1 overexpression in osteoprogenitor cells promoted matrix mineralization of osteoblast differentiation but did not promote matrix maturation. DKK1 increased Ca+ influx and activation of the Ca+/calmodulin-dependent protein kinase II Alpha (CAMK2A)-cAMP response element-binding protein 1 (CREB1) and increased translocation of p-CREB1 into the nucleus. In contrast, stable DKK1 knockdown in human osteosarcoma cell line SaOS2 exhibited reduced nuclear translocation of p-CREB1 and matrix mineralization. Overall, we suggest that manipulating DKK1 regulates the matrix mineralization of osteoblasts by Ca+-CAMK2A-CREB1, and DKK1 is a crucial gene for bone mineralization of osteoblasts.

Re-Identification on Korean Penicillium Sequences in GenBank Collected by Software GenMine

  • Chang Wan Seo;Sung Hyun Kim;Young Woon Lim;Myung Soo Park
    • Mycobiology
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    • v.50 no.4
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    • pp.231-237
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    • 2022
  • Penicillium species have been actively studied in various fields, and many new and unrecorded species continue to be reported in Korea. Moreover, unidentified and misidentified Korean Penicillium species still exist in GenBank. Therefore, it is necessary to revise the Korean Penicillium inventory based on accurate identification. We collected Korean Penicillium nucleotide sequence records from GenBank using the newly developed software, GenMine, and re-identified Korean Penicillium based on the maximum likelihood trees. A total of 1681 Korean Penicillium GenBank nucleotide sequence records were collected from GenBank. In these records, 1208 strains with four major genes (Internal Transcribed Spacer rDNA region, b-tubulin, Calmodulin and RNA polymerase II) were selected for Penicillium reidentification. Among 1208 strains, 927 were identified, 82 were identified as other genera, the rest remained undetermined due to low phylogenetic resolution. Identified strains consisted of 206 Penicillium species, including 156 recorded species and 50 new species candidates. However, 37 species recorded in the national list of species in Korea were not found in GenBank. Further studies on the presence or absence of these species are required through literature investigation, additional sampling, and sequencing. Our study can be the basis for updating the Korean Penicillium inventory.

Characterization of Newly Recorded Talaromyces veerkampii Isolated from Field Soil in Korea based on Morphology and Multigene Sequence Analysis

  • Mahesh Adhikari;Hyun Seung Kim;Hyo Bin Park;Ki Young Kim;In Kyu Lee;Eun Jeong Byeon;Ji Min Woo;Hyang Burm Lee;Youn Su Lee
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.347-355
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    • 2022
  • A fungal isolate belonging to the phylum Ascomycota was isolated and identified as Talaromyces veerkampii in 2017 during a survey of fungal diversity in field soils in Korea. This fungal isolate was identified and described based on macro- and micromorphological and molecular characterization. The identification was also based on partial 18S-ITS1-5.8S-ITS2-28S rDNA and calmodulin (CaM)-encoding gene sequencing data. Talaromyces veerkampii has not been previously reported in Korea. Thus, we report here a newly discovered species from soil in Korea along with its morphological and molecular characteristics.

Isolation and Identification of an Unreported Fungal Species in Korea and Novel Ice Nucleation Active Fungus: Fusarium diversisporum

  • Diane Avalos-Ruiz;Gwang-Jae Lim;Seong-Keun Lim;Leonid N. Ten;In-Kyu Kang;Seung-Yeol Lee;Hee-Young Jung
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.255-262
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    • 2022
  • In this study, the fungal strain KNUF-21-F39 was isolated from a declined apple tree (Malus domestica) in the Chungcheongbuk province in Korea. The strain KNUF-21-F39 presented a slow growth rate and a variety of macroconidia shapes and sizes ranging from ovoid to fusoid and 1- to 5-septate, primarily showing 3- and 4-septate, with "S" -shaped macroconidia rarely observed. The strain was identified based on morphological characteristics along with phylogenetic analysis performed using the internal transcribed spacer region (ITS) and partial sequences of translation elongation factor 1-α (tef1), RNA polymerase largest subunit (rpb1), and calmodulin (cal) genes. The fungal strain KNUF-21-F39 was identified as Fusarium diversisporum, which has not been previously reported in Korea. The ice nucleation activity (INA) of the strain was also evaluated, identifying the strain as positive for INA. This is the first report characterizing F. diversisporum as an IN-active fungal species.