• Title/Summary/Keyword: cytoplasmic localization

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Ultrastructural and Cytochemical Studies on the Endometrial Stromal Cells of Guinea Pig during Estrous Cycle (발정주기(發情週期)에 따른 Guinea Pig의 자궁내막(子宮內膜) 기질세포(基質細胞)의 미세구조(微細構造)및 세포화학적(細胞化學的) 연구(硏究))

  • Deung, Young-K.;You, Kwan-H.;Choi, Choon-K.;Lee, Choon-K.;Chung, Ho-S.
    • Applied Microscopy
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    • v.12 no.1
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    • pp.57-68
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    • 1982
  • Cyclical changes in the fine structures of endometrial stroma of guinea pig during the estrous cycle were studied by electron microscopy. Cytochemical studies were made in order to investigate the ultrastructural localization of the acid phosphatase,alkaline phosphatase and ATPase in these cells. The results obtained are as follows: 1. During estrus collagen fibers were most abundant in the stroma. The stromal cells showed increases in the number of several cytoplasmic organelles, especially the rough endoplasmic reticulum was significantly increased and the structures were greatly differentiated. 2. Many cytoplasmic processes and cell debris have been distributed in the stroma of metestrus. The distributions were increased and degenerated mitochondria were observed during diestrus. 3. Cytochemical studies indicated that during metestrus and diestrus acid phosphatase activities were localized in the degenerating collagen fibers. Alkaline phosphatase activities were weak in the collegen fibers during proestrus and estrus which intense activities were localized around the cell membrane during metestrus and diestrus. ATPase activities were present on the cell membrane and intercellular space of stromal cell during proestrus and estrus.

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Screening and Analysis for cTPx II-Interacting Protein Using Yeast Wo-hybrid System (Yeast Two-hybrid System을 이용한 cTPx II 결합단백질 탐색 및 분석)

  • Kim. Il-Han;Oh, Young-Mee;Cha, Mee-Kyung
    • The Journal of Natural Sciences
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    • v.15 no.1
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    • pp.79-88
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    • 2005
  • There are five isoforms of thiol peroxidase in yeast. Each isoform was named after its subcellular localization such as cytoplasmic TPx I, cTPx II, cTPx III, mitochondrial TPx (mTPx), and nuclear TPx (nTPx). Recently, we reported that unlike other TPx null mutants, cTPx IInull mutant showed a slow-growth phenotype. This observation suggests that cTPx II might be involved in yeast cell growth. In this study, for a first step toward to investigate the physiological function of cTPx II in yeast, we have identified a novel interaction between cTPx II and various proteins by using the yeast two-hybrid system.

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Localization of Cyclin B and Erk1/2 in Ovine Oocytes and MPF and MAPK Activities in Cytoplast and Karyoplast following Enucleation

  • Lee, Joon-Hee;Campbell, Keith H.S.
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.407-414
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    • 2011
  • The development of embryos reconstructed by somatic cell nuclear transfer (SCNT) is dependent upon numerous factors. Central to development is the quality and developmental competence of the recipient cytoplast and the type of the donor nucleus. Typically metaphase of the second meiotic division (MII) has become the cytoplast of choice. Production of a cytoplast requires removal of the recipient genetic material, however, it may remove proteins which are essential for development or reduce the levels of cytoplasmic proteins to influence subsequent reprogramming of the donor nucleus. In this study, enucleation at MII did not affect the activities of either MPF or MAPK kinases. Immunocytochemical staining showed that both Cyclin B1 (MPF) and Erk1/2 (MAPK) were associated with the meiotic spindle of AI/TI oocytes with little staining in the cytoplasm, however, at MII association of both proteins with the spindle had reduced and a greater degree of cytoplasmic distribution was observed. The analysis of oocyte proteins removed during enucleation is a difficult approach to the identification of factors which may be depleted in the cytoplast. This is primarily due to the large numbers of aspirated karyoplasts which would be required for the analysis.

Phosphorylation of p53 at threonine 155 is required for Jab1-mediated nuclear export of p53

  • Lee, Eun-Woo;Oh, Wonkyung;Song, Hosung Paul;Kim, Won Kon
    • BMB Reports
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    • v.50 no.7
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    • pp.373-378
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    • 2017
  • The Jun activation-domain binding protein 1 (Jab1) induces p53 nuclear export and cytoplasmic degradation, but the underlying mechanism is poorly understood. Here, we show that phosphorylation at the threonine 155 residue is essential for Jab1-mediated p53 nuclear export. Jab1 stimulated phosphorylation of p53 at T155 was inhibited by curcumin, an inhibitor of COP9 signalosome (CSN)-associated kinases. The T155E mutant, which mimics phosphorylated p53, exhibited spontaneous cytoplasmic localization in the absence of Jab1. This process was prevented by leptinomycin B (LMB), but not by curcumin. The substitution of threonine 155 for valine (T155V) abrogated Jab1-mediated p53 nuclear export, indicating that phosphorylation at this site is essential for Jab1-mediated regulation of p53. Although T155E can be localized in the cytoplasm in the absence of Mdm2, the translocation of T155E was significantly enhanced by ectopic Hdm2 expression. Our data suggests that Jab1-mediated phosphorylation of p53 at Thr155 residue mediates nuclear export of p53.

Localization of Transferrin mRNA in Rat by DNA/RNA Hybridization (DNA/RNA Hybridization에 의한 흰쥐의 Transferrin mRNA 분포에 관한 연구)

  • Kim, Se-Eun;Kim, Sun-Yeou;Park, Mi-Jung;Song, Jin-Ho;Lee, Eun-Bang;Lee, Heun-Pa;Kim, Young-Choong
    • YAKHAK HOEJI
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    • v.33 no.5
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    • pp.300-307
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    • 1989
  • Expression of transferrin gene in various organs of rat was studied using rat transferrin cDNA. The hybridization method of $[^{35}S]-labeled$ transferrin cDNA with transferrin mRNA in cytoplasmic preparations was used to measure the level of transferrin mRNA. The rat from 15-day old fetus to 21-day old postnatal were employed as an animal model. In the liver, the level of transferrin mRNA increased with increasing age. However, the level of transferrin mRNA in brain was significantly lower than that in liver and the level did not increase with age.

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Enzymological Localization of Carbon Monoxide Dehydrogenases in Pseudomonas carboxydovorans and Acinetobacter sp.1 (Pseudomonas carboxydovorans와 Acinetobacter sp.1의 일산화탄소 산화효소의 세포내 분포에 대한 효소학적 연구)

  • 김시욱;김영민
    • Korean Journal of Microbiology
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    • v.24 no.3
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    • pp.270-275
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    • 1986
  • The localization of carbon monoxide dehydrogenases (CO-DHs) in Pseudomonas carvoxydovorans and Acinetobacter sp.1 was examined by comparison of the distribution of CO-oxidizing activity between soluble and particulate fractions obtained after disruption of CO-grown cells by sonic oscillation and of spheroplasts by osmotic shock. When the cells were broken by sonic oscillation, most of the CO-DH activity was recovered from soluble fractions. However, disryption by osmotic lysis of spheroplasts revealed that the enzyme activity is present in the cell membrane. The results indicated the CO-DHs in these cells are loosely attached to the cytoplasmic membrane.

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재조합 Saccharomyces cerevisiae에서 Inulinase와 Invertase의 발현과 분비에 미치는 배양조건의 영향

  • 남수완;신동하;김연희
    • Microbiology and Biotechnology Letters
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    • v.25 no.3
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    • pp.258-265
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    • 1997
  • The effects of medium pH and culture temperature on the expression and secretion of inulinase and invertase were investigated with recombinant Saccharomyces cerevisiae cells. These cells were obtained by transformation of 2$\mu$-based plasmids pYI10 and pYS10 which contain Kluyveromyces marxianus inulinase gene (INU1A) and S. cerevisiae invertase gene (SUC2), respectively, in the downstream of GAL1 promoter. The expression level and localization of inulinase and invertase were not affected significantly by the initial medium pH: secretion efficiencies of inulinase and invertase into the medium were about 90% and 60%, respectively, in the pH ranges of 4.0 to 6.5. However, the expression and secretion of both enzymes were strongly dependent on the culture temperature. The highest expression (7.7 units/mL) and secretion (6.7 units/mL) of inulinase were observed at 28$\circ$C and 30$\circ$C. As a consequence of decreased localization of inulinase in the periplasmic space, the secretion efficiency increased from 68% at 20$\circ$C, to 95% at 35$\circ$C,. The total expression level and secretion efficiency of invertase increased from 19 units/mL and 55% at 20$\circ$C to 25 units/mL and 68% at 35$\circ$C, respectively. Irrespective of the culture temperature, the invertase activity in the cellular fraction (periplasmic space and cytoplasmic fractions) was kept constant at around 33-45%.

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Increased Expression of Epithelial Cell Adhesion Molecule (EpCAM) in Rat Hepatic Tumors Induced by Diethylnitrosamine

  • Kang, Jin Seok
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3627-3630
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    • 2012
  • The epithelial cell adhesion molecule (EpCAM) is a pan-epithelial differentiation antigen that is expressed on almost all carcinomas. However, a role in rat liver carcinogenesis has never been reported previously. Thus, its expression was investigated herein in rat liver tumors induced by diethylnitrosamine (DEN). Twenty male 5-week-old F344 rats were used in this experiment. Mini-osmotic pumps containing doses of 47.5 mg of DEN were inserted into the abdominal cavity of each animal to initiate liver carcinogenesis. All animals were sacrificed at 26 weeks after DEN treatment. At necropsy, hepatic masses were processed for histopathological examination, which revealed forty-four hepatocellular adenomas (HCAs) and twenty hepatocellular carcinomas (HCC). Tumors were immunohistochemically analyzed for EpCAM, proliferating cell nuclear antigen (PCNA) and co-localization of the two. EpCAM expression was mainly detected in hepatic tumor cells, showing a cytoplasmic staining pattern. However, expression was also slightly observed in normally-appearing surrounding hepatic cells. PCNA expression was highly detected in tumor cells, showing nuclear staining. Double staining of EpCAM and PCNA in tumors showed many cells with co-localization. Taken together, EpCAM and PCNA expression were increased in DEN-induced tumors and many tumor cells showed co-expression. It is suggested that EpCAM may increase during DEN-induced tumors, possibly associated with cell proliferation.

HDAC4 Regulates Muscle Fiber Type-Specific Gene Expression Programs

  • Cohen, Todd J.;Choi, Moon-Chang;Kapur, Meghan;Lira, Vitor A.;Yan, Zhen;Yao, Tso-Pang
    • Molecules and Cells
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    • v.38 no.4
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    • pp.343-348
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    • 2015
  • Fiber type-specific programs controlled by the transcription factor MEF2 dictate muscle functionality. Here, we show that HDAC4, a potent MEF2 inhibitor, is predominantly localized to the nuclei in fast/glycolytic fibers in contrast to the sarcoplasm in slow/oxidative fibers. The cytoplasmic localization is associated with HDAC4 hyper-phosphorylation in slow/oxidative-fibers. Genetic reprogramming of fast/glycolytic fibers to oxidative fibers by active CaMKII or calcineurin leads to increased HDAC4 phosphorylation, HDAC4 nuclear export, and an increase in markers associated with oxidative fibers. Indeed, HDAC4 represses the MEF2-dependent, PGC-$1{\alpha}$-mediated oxidative metabolic gene program. Thus differential phosphorylation and localization of HDAC4 contributes to establishing fiber type-specific transcriptional programs.

Visualization of Phytophthora palmivora Infection in Oil Palm Leaflets with Fluorescent Proteins and Cell Viability Markers

  • Ochoa, Juan C.;Herrera, Mariana;Navia, Monica;Romero, Hernan Mauricio
    • The Plant Pathology Journal
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    • v.35 no.1
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    • pp.19-31
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    • 2019
  • Bud rot (BR) is the most devastating disease affecting oil palm (Elaeis guineensis) crops in Colombia. Its causal agent, Phytophthora palmivora, initiates the infection in immature oil palm leaflets producing necrotic lesions, followed by colonization of opportunistic necrotrophs, which increases disease damage. To improve the characterization of the disease, we transformed P. palmivora using Agrobacterium tumefaciens-mediated transformation (ATMT) to include the fluorescent proteins CFP-SKL (peroxisomal localization), eGFP and mRFP1 (cytoplasmic localization). The stability of some transformants was confirmed by Southern blot analysis and single zoospore cultures; additionally, virulence and in vitro growth were compared to the wild-type isolate to select transformants with the greatest resemblance to the WT isolate. GFP-tagged P. palmivora was useful to identify all of the infective structures that are commonly formed by hemibiotrophic oomycetes, including apoplastic colonization and haustorium formation. Finally, we detected cell death responses associated with immature oil palm tissues that showed reduced susceptibility to P. palmivora infection, indicating that these tissues could exhibit age-related resistance. The aim of this research is to improve the characterization of the initial disease stages and generate cell biology tools that may be useful for developing methodologies for early identification of oil palm materials resistant or susceptible to BR.