• Title/Summary/Keyword: gene localization

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Insulin-Like Growth Factor-I Induces Androgen Receptor Activation in Differentiating C2C12 Skeletal Muscle Cells

  • Kim, Hye Jin;Lee, Won Jun
    • Molecules and Cells
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    • v.28 no.3
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    • pp.189-194
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    • 2009
  • The modulating effect of IGF-I on the regulation of AR gene expression and activation in skeletal muscle cells remains poorly understood. In this study, the effects of IGF-I treatment on AR induction and activation in the absence of AR ligands were examined. Differentiating C2C12 cells were treated with different concentrations (0-250 ng/ml) of IGF-I or for various periods of time (0-60 min) of 250 ng/ml IGF-I. Treatment of C2C12 cells with IGF-I resulted in a dose- and time-dependent increase in total AR and phosphorylated AR (Ser 213). IGF-I treatment also led to significantly increased AR mRNA expression when compared with the control. The levels of skeletal ${\alpha}-actin$ and myogenin mRNA, known target genes of AR, were also significantly upregulated after 5 or 10 min of treatment with IGF-I. Confocal images revealed that IGF-I stimulated nuclear localization of AR in the absence of ligands. In addition, an electrophoretic mobility shift assay indicated that IGF-I stimulated the AR DNA binding activity in a time-dependent manner. The present results suggest that IGF-I stimulates the expression and activation of AR by ligand-independent mechanism in differentiating C2C12 mouse skeletal muscle cells.

Expression of EuNOD-ARP1 Encoding Auxin-repressed Protein Homolog Is Upregulated by Auxin and Localized to the Fixation Zone in Root Nodules of Elaeagnus umbellata

  • Kim, Ho Bang;Lee, Hyoungseok;Oh, Chang Jae;Lee, Nam Houn;An, Chung Sun
    • Molecules and Cells
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    • v.23 no.1
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    • pp.115-121
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    • 2007
  • Root nodule formation is controlled by plant hormones such as auxin. Auxin-repressed protein (ARP) genes have been identified in various plant species but their functions are not clear. We have isolated a full-length cDNA clone (EuNOD-ARP1) showing high sequence homology to previously identified ARP genes from root nodules of Elaeagnus umbellata. Genomic Southern hybridization showed that there are at least four ARP-related genes in the genome of E. umbellata. The cDNA clone encodes a polypeptide of 120 amino acid residues with no signal peptide or organelle-targeting signals, indicating that it is a cytosolic protein. Its cytosolic location was confirmed using Arabidopsis protoplasts expressing a EuNOD-ARP1:smGFP fusion protein. Northern hybridization showed that EuNOD-ARP1 expression was higher in root nodules than in leaves or uninoculated roots. Unlike the ARP genes of strawberry and black locust, which are negatively regulated by exogenous auxin, EuNOD-ARP1 expression is induced by auxin in leaf tissue of E. umbellata. In situ hybridization revealed that EuNOD-ARP1 is mainly expressed in the fixation zone of root nodules.

Cloning, Sequencing and Characterization of Mitochondrial Control Region of the Domestic Silkwom, Bombyx mori

  • Lee, Jin-Sung;Kim, Ki-Hwan;Hoe, Hyang-Sook;Park, Jae-Heung;Kang, Seok-Woo;Lee, Sang-Han;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.87-89
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    • 2001
  • The nucleotide sequence of the domestic silkworm (Bombyx mori) mitochondrial (mt) control region and its flanking genes was determined from PCR clones. The control region of the silkworm mt genome was located between the small ribosomal RNA gene and transfer RN $A^{Met}$. This 499 bp control region hale 95.4% A+T content. Extensive comparative analysis studies performed with similar control region of other insect genomes could not reveal a highly conserved region containing conserved motifs of animal mito-chondrial genome. The remarkable feature that found in this control region was the presence of tandem motifs containing nine repetitive sequences. The potential usefulness of this motif sequences for Bombyx species or their taxonomically related species is enhanced by its unique localization in the maternally inheritance mitochondrial molecule.e.

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Immunohistochemical studies on the distribution of neuropeptides in the tongue of Korean native goat (한국재래산양 혀에 분포하는 신경전달물질에 관한 면역조직화학적 연구)

  • Lee, Heungshik S.;Lee, In-se;Kang, Tae-cheon
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.265-276
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    • 1996
  • This study was performed to identify the localization of several neuropeptides; calcitonin gene-related peptide(CGRP), substance P(SP), vasoactive intestinal polypeptide(VIP), neuropeptide Y(NPY), serotonin(5-HT) and neurotensin in the tongue of Korean native goat(Capra hircus) by immunohistochemical method. The results were summarized as follows: CGRP- and SP-immunoreactive fibers were observed as moderate immunoreactivity at the subepithelial plexus and subgemmal fibers in lamina propria of lingual papillae, but not seen in intragemmal, intergemmal, perigemmal fibers as well as in the supporting, basal and taste cells. Fibers around the acinus of the von Ebner's gland and blood vessels showed weak immunoreactivities against CGRP and SP. In the intrinsic ganglion cells, CGRP- and SP-immunoreactivities were not observed. The distribution patterns of VIP- and NPY-immunoreactive fibers were similar to CGRP-or SP-immunoreactive fibers, but their immunoreactivities were stronger than those of CGRP- or SP immunoreactive fibers. The immunoreactivities to VIP or NPY were seen in the intrinsic ganglion. Only a few serotonin immunoreactive fibers were seen in some filiform or fungiform papillae. Neurotensin immunoreactivity was not observed in the tongue of Korean native goat.

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Characterization of the cellular localization of C4orf34 as a novel endoplasmic reticulum resident protein

  • Jun, Mi-Hee;Jun, Young-Wu;Kim, Kun-Hyung;Lee, Jin-A;Jang, Deok-Jin
    • BMB Reports
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    • v.47 no.10
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    • pp.563-568
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    • 2014
  • Human genome projects have enabled whole genome mapping and improved our understanding of the genes in humans. However, many unknown genes remain to be functionally characterized. In this study, we characterized human chromosome 4 open reading frame 34 gene (hC4orf34). hC4orf34 was highly conserved from invertebrate to mammalian cells and ubiquitously expressed in the organs of mice, including the heart and brain. Interestingly, hC4orf34 is a novel ER-resident, type I transmembrane protein. Mutant analysis showed that the transmembrane domain (TMD) of hC4orf34 was involved in ER retention. Overall, our results indicate that hC4orf34 is an ER-resident type I transmembrane protein, and might play a role in ER functions including $Ca^{2+}$ homeostasis and ER stress.

Molecular characterization and immunohistochemical localization of a mitogen-activated protein kinase, Accp38b, from Apis cerana cerana

  • Zhang, Liang;Meng, Fei;Li, Yuzhen;Kang, Mingjiang;Guo, Xingqi;Xu, Baohua
    • BMB Reports
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    • v.45 no.5
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    • pp.293-298
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    • 2012
  • The p38 mitogen-activated protein kinase (MAPK) is involved in various processes, including stress responses, development, and differentiation. However, little information on p38 MAPK in insects is available. In this study, a p38 MAPK gene, $Accp38b$, was isolated from $Apis$ $cerana$ $cerana$ and characterized. The quantitative real-time PCR (Q-PCR) analysis revealed that $Accp38b$ was induced by multiple stressors. Notably, the expression of $Accp38b$ was relatively higher in the pupae phase than in other developmental phases. During the pupae phase, Accp38b expression was higher in the thorax than in the head and abdomen and higher in the fat body than in the muscle and midgut. Immunohistochemisty showed significant positive staining of Accp38b in sections from the brain, eyes, fat body, and midgut of $A.$ $cerana$ $cerana$. These results suggest that Accp38b may play a crucial role in stress responses and have multiple aspects function during development.

Heterogeneous overexpression of Oryza sativa salt induced RING Finger protein OsSIRF1 positively regulates salt and osmotic stress in transgenic Arabidopsis

  • Chapagain, Sandeep;Jang, Cheol Seong
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.150-150
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    • 2017
  • Plants suffer from various abiotic stresses among them; soil salinity is one of major adverse factor in declining agricultural productivity. So, development of salt stress tolerance crops have potential role to increase crop production. The RING finger proteins are known to play crucial roles in abiotic stress environment to plants. In this study, we identified one Salt-responsive Really${\underline{I}nteresting}$ ${\underline{n}ew}$ ${\underline{g}ene}$ (RING) E3 ubiquitin ligase gene OsSIRF1 from rice root tissues during salt stress and studied its molecular function. Expression of OsSIRF1 was induced under various abiotic stress conditions, including salt, heat, drought, and ABA. Result of an in vitro ubiquitination assay clearly showed that OsSIRF1 Possess an E3 ligase activity. Moreover, OsSIRF1 was found to be localized to the nucleus within the cell. Heterogeneous overexpression of OsSIRF1 in Arabidopsis improved seed germination and increased root length under salt and Manitol stress conditions. Taking together, these results suggested that OsSIRF1 may be associated with plant responses to abiotic stressors and positively regulates salt and osmotic stress environment.

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C-terminally mutated tubby protein accumulates in aggresomes

  • Kim, Sunshin;Sung, Ho Jin;Lee, Ji Won;Kim, Yun Hee;Oh, Yong-Seok;Yoon, Kyong-Ah;Heo, Kyun;Suh, Pann-Ghill
    • BMB Reports
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    • v.50 no.1
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    • pp.37-42
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    • 2017
  • The tubby protein (Tub), a putative transcription factor, plays important roles in the maintenance and function of neuronal cells. A splicing defect-causing mutation in the 3'-end of the tubby gene, which is predicted to disrupt the carboxy-terminal region of the Tub protein, causes maturity-onset obesity, blindness, and deafness in mice. Although this pathological Tub mutation leads to a loss of function, the precise mechanism has not yet been investigated. Here, we found that the mutant Tub proteins were mostly localized to puncta found in the perinuclear region and that the C-terminus was important for its solubility. Immunocytochemical analysis revealed that puncta of mutant Tub co-localized with the aggresome. Moreover, whereas wild-type Tub was translocated to the nucleus by extracellular signaling, the mutant forms failed to undergo such translocation. Taken together, our results suggest that the malfunctions of the Tub mutant are caused by its misfolding and subsequent localization to aggresomes.

Molecular and Cellular Studies of Seed Storage Proteins from Rice and Wheat

  • Kim, Woo-Taek
    • Applied Biological Chemistry
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    • v.32 no.1
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    • pp.64-72
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    • 1989
  • Near full length cDNA clones encoding the rice seed storage protein, prolamine, were isolated and divided into two homology classes based on cross-hybridization and DNA sequencing analysis. These cDNA clones contain a single open reading frame encoding a putative rice prolamine precursor(M.W.=17,200) possessing atypical 14 amino acid signal peptide. Clones of these two homology classes diverge mainly by insertions/deletions of short nucleotide stretches and point mutations. The deduced primary structures of both types of prolamine polypeptides are devoid of any major tandem repetitive sequences, a feature prevalent in other cereal prolamines. No significant homology teas detected between the rice prolamine and other cereal prolamines, indicating that the rice gene evolved from a different ancestor that gave rise to other cereal prolamine genes. Developing wheat and rice endosperms were examined using ultrathin sections prepared from tissues harvested at various days after flowering. By immunocytochemical localization techniques, wheat prolamines are localized within vesicles from Golgi apparatus and in homogeneous regions of protein bodies. The involvement of the goli apparatus in the packaging of wheat prolamines into protein bodies indicates a pathway which differs from the mode of other cereal prolamines and resembles the mechanism employed for the storage of rice glutelin and legume globulins.

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Molecular cloning and characterization of a soybean GmMBY184 induced by abiotic stresses

  • Chung, Eun-Sook;Kim, Koung-Mee;Lee, Jai-Heon
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.175-181
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    • 2012
  • Drought and high salinity stresses often imposes adverse effects on crop yield. MYB transcription factors have been shown to be an important regulator in defense responses to these environmental stresses. In this study, we have cloned and characterized a soybean gene GmMYB184 (Glycine max MYB transcription factor 184). Deduced amino acid sequences of GmMYB184 show highest homology with that from Vitis vinifera legume plant (75%). Different expression patterns of GmMYB184 mRNA were observed subjected to drought, cold, high salinity stress and abscisic acid treatment, suggesting its role in the signaling events in the osmotic stress-related defense response. Subcellular localization studies demonstrated that the GFP-GmMYB184 fusion protein was localized in the nucleus. Using the yeast assay system, the C-terminal region of GmMYB184 was found to be essential for the transactivation activity. These results indicate that the GmMYB184 may play a role in abiotic stress tolerance in plant.