• Title/Summary/Keyword: cytoplasmic localization

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Vinyl-Stilbene Inhibits Human Norovirus RNA Replication by Activating Heat-Shock Factor-1

  • Lee, Ahrim;Sung, Jieun;Harmalkar, Dipesh S.;Kang, Hyeseul;Lee, Hwayoung;Lee, Kyeong;Lee, Choongho
    • Biomolecules & Therapeutics
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    • v.30 no.1
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    • pp.64-71
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    • 2022
  • Norovirus (NV) is the most common cause of viral gastroenteritis, with the potential to develop into a fatal disease in those who are immuno-compromised, and effective vaccines and treatments are still non-existent. In this study, we aimed to elucidate the molecular mechanism of the previously identified NV replication inhibitor utilizing a vinyl-stilbene backbone, AC-1858. First, we confirmed the inhibition of the NV RNA replication by a structural analog of AC-1858, AC-2288 with its exclusive cytoplasmic sub-cellular localization. We further validated the induction of one specific host factor, the phosphorylated form of heat shock factor (HSF)-1, and its increased nuclear localization by AC-1858 treatment. Finally, we verified the positive and negative impact of the siRNA-mediated downregulation and lentivirus-mediated overexpression of HSF-1 on NV RNA replication. In conclusion, these data suggest the restrictive role of the host factor HSF-1 in overall viral RNA genome replication during the NV life cycle.

Secondary Structure, 1H, 13C and 15N Resonance Assignments and Molecular Interactions of the Dishevelled DIX Domain

  • Capelluto, Daniel G.S.;Overduin, Michael
    • BMB Reports
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    • v.38 no.2
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    • pp.243-247
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    • 2005
  • Dishevelled (Dvl) is a positive regulator of the canonical Wnt signaling pathway, which regulates the levels of $\beta$-catenin. The $\beta$-catenin oncoprotein depends upon the association of Dvl and Axin proteins through their DIX domains, and its accumulation directs the expression of specific developmental-related genes at the nucleus. Here, the $^1H$, $^{13}C$, and $^{15}N$ resonances of the human Dishevelled 2 DIX domain are assigned using heteronuclear nuclear magnetic resonance (NMR) spectroscopy. In addition, helical and extended elements are identified based on the NMR data. The results establish a structural context for characterizing the actin and phospholipid interactions and binding sites of this novel domain, and provide insights into its role in protein localization to stress fibers and cytoplasmic vesicles during Wnt signaling.

Immunofluorescence Localization of Schizosaccharomjyces pombe $cdc103^{+}$ Gene Product

  • Kim, Hyong-Bai
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.248-254
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    • 1996
  • $cdc103^+$ gene in Schizosaccharomyces pombe which is similar to the CDC3 gene in Saccharomyces cerevisiae was cloned and sequenced. Comparison of the predicted amino acid sequences of $cdc103^+$ and CDC3 revealed that they share significant similarity (43% identity and 56% identity or similarity) to each other. The gene product of CDC3 in S. cerevisiae is known to be a highly ordered ring of filaments that lies just inside the cytoplasmic membrane in the region of the mother-bud neck. In order to characterize the gene product of $cdc103^+$ in Schizosaccharomyces pombe, fusion proteins were used to generate the polyclonal antibodies specific for the gene product (cdc103p). In immunofluorescence experiments, these antibodies decorate the region of the septum formation as a double ring structure late in the cell division cycle.

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Comparison of Exon-boundary Old and Young Domains during Metazoan Evolution

  • Lee, Byung-Wook
    • Genomics & Informatics
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    • v.7 no.2
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    • pp.131-135
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    • 2009
  • Domains are the building blocks of proteins. Exon shuffling is an important mechanism accounting for combination of a limited repertoire of protein domains in the evolution of multicellular species. A relative excess of domains encoded by symmetric exons in metazoan phyla has been presented as evidence of exon shuffling, and symmetric domains can be divided into old and new domains by determining the ages of the domains. In this report, we compare the spread, versatility, and subcellular localization of old and new domains by analyzing eight metazoan genomes and their respective annotated proteomes. We found that new domains have been expanding as multicellular organisms evolved, and this expansion was principally because of increases in class 1-1 domains amongst several classes of domain families. We also found that younger domains have been expanding in membranes and secreted proteins along with multi-cellular organism evolution. In contrast, old domains are located mainly in nuclear and cytoplasmic proteins. We conclude that the increasing mobility and versatility of new domains, in contrast to old domains, plays a significant role in metazoan evolution, facilitating the creation of secreted and transmembrane multidomain proteins unique to metazoa.

Cell-type specific expression of vanilloid receptor 1 in the taste cells of rat circumvallate papillae

  • Moon, Young-Wha;Han, Ji-Won;Kang, Wha-Sun
    • Animal cells and systems
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    • v.15 no.3
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    • pp.197-202
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    • 2011
  • The present study demonstrates the first-ever characterization of cell types that express the vanilloid receptor 1 (VR1) in the taste buds of rat circumvallate papillae. We performed electron microscopy to identify the subcellular location. The VR1 immunoreactivity was associated with the endoplasmic reticulum, cytoplasmic vesicles, and plasma membrane of taste cells. These results demonstrate the localization of the VR1 in membranous structures of the taste cells. We used double immunofluorescence histochemistry with taste cell type-specific markers to identify the cell types that express the VR1. The VR1 was detected in all functional taste cell types (Type I, Type II, and Type III cells). Together, our data suggest that the VR1 might play different roles according to the cell types within a taste bud.

Expression of Bone Morphogenetic Protein-2 and Histological Differentiation of Oral Squamous Cell Carcinomas

  • Hamasni, Fatme Mouchref;El Hajj, Fady
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.12
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    • pp.5243-5245
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    • 2016
  • Background and Objective : Bone morphogenic protein-2 (BMP-2) plays an essential role in mesenchymal cell differentiation into osteoblasts، through many intracellular pathways which may also be active in tumors. Invasive oral squamous cell carcinomas account for more than 90% of head and neck malignancies in many cancer registries. They are classified into three types according to epithelial cell differentiation. The present study aimed to identify any relation between BMP-2 expression and tumor histology. Materials and methods: BMP-2 expression was compared immunohistochemically among 30 cases (19 male and 11 female, mean age 48 years) of oral squamous cell carcinoma, Division was into 3 groups (each containing 10 cases) according to the histological grade. Results: No significant correlation between BMP-2 expression and histological grade was observed. Changes in localization and cytoplasmic staining were also not apparent. Conclusion: From the results of this study BMP-2 does not appear to have any application as a prognostic marker for oral squamous cell carcinomas.

The innate immune response transcription factor Bombyx mori Relish1 induces high-level antimicrobial peptides in silkworm

  • Kim, Seong-Wan;Kim, Seong-Ryul;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.37 no.2
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    • pp.49-54
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    • 2018
  • To artificially enhance antimicrobial peptide expression in Bombyx mori, we constructed genetically engineered silkworms overexpressing Rel family transcription factor. The truncated BmRelish1 (BmRelish1t) gene contained a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acid (AHAA)-rich region, and death domain (DD), but no ankyrin-repeat (ANK) domain. The BmRelish1t gene was controlled by B. mori cytoplasmic actin 3 promoter in the PiggyBac transposon vector. Chromosome analysis of G1 generations of a transgenic silkworm with EGFP expression confirmed stable insertion of BmRelish1t. BmRelish1t gene overexpression in transgenic silkworms resulted in higher mRNA expression levels of B. mori antimicrobial peptides such as lebocin(~20.5-fold), moricin(~8.7-fold), and nuecin(~17.4-fold) than those in normal silkworms.

Specific Localization of DNMT1 in Mouse and Bovine Preimplantation Embryos

  • Y.M.Chang;Min, K.S.;Yoon, J.T.;M.G.Pang;Chung, Y.C.;Kim, C.K.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.81-81
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    • 2003
  • DNA methylation is a covalent modification of DNA that can modulate gene expression and is now recognized as a major component of the epigenome. During evolution, the dinucleotide CpG has been progressively eliminated from the genome of higher eukaryotes and is present at only 5% to 10% of its predicted frequency. Approxymately 80% of the remaining CpG sites contain methylated cytosines in most vertebrates and they are distributed in a pattern that is unique in each tissue and is inversely correlated with gene expression. The pattern of methylation is faithfully maintained during cell division by the enzyme Dnmt1, the maintenance DNA methyltransferase, which catalyzes the transfer of a methyl group from S-adenosyl-methionine to the 5'-position of the cytosine ring. We have been identified bovine Dnmt1 cDNA full-length recently (AY173048) Little is known on the functions of Dnmt1 in bovine preimplantation embryos. Thus, we analyzed the specific pattern of Dnmt1 in in vitro derived/nuclear transfer bovine and in vivo derived mouse embryos to monitor the epigenetic reprogramming process. We investigated these process by using indirect immunofluresence with an antibody to Dnmt1. According to other studies, Dnmt1 accumulates in nuclei of early growing oocytes but is sequestered in the cytoplasm of mature oocytes. In 2-cell and 4-cell embryos, Dnmt1 is cytoplasmic, but at the 8-cell stage, it is present only in the nucleus. By the blastocyst stage, Dnmt1o is again found only in the cytoplasm. Thus, nuclear localization of Dnmt1o in preimplantation embryos is limited to the 8-cell stages After implantation, Dnmt1 is localized in the nucleus in mouse. However, we have found different patterns of Dnmt1 nuclear localization. Though we used the common antibody, immune-localization data revealed that Dnmt1 antibody have been detected at the nucleus in 1-cell to blastocyst embryos. Therefore, maybe we think that the functions of Dnmt1 between bovine and mice are different. In order to Identify the mechanisms that regulate DNA methylation in bovine preimplantation embryo, we have plans on using bovine oocyte and somatic specific Dnmt1 antibodies.

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Fine Needle Aspiration Cytology of Glycogen-Rich Clear Cell Carcinoma of the Breast - A Report of 2 Cases - (유방의 당원성 투명세포암종의 세침흡인 세포학적 소견 - 2예 보고 -)

  • Kim, Wan-Seop;Lee, Won-Mi;Hong, Eun-Kyung;Park, Moon-Hyang;Lee, Jung-Dal
    • The Korean Journal of Cytopathology
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    • v.9 no.2
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    • pp.213-219
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    • 1998
  • Glycogen-rich clear ceil carcinoma of the breast is an unusual variant of carcinoma with a recorded incidence of $1.4{\sim}3%$ of breast carcinomas. The cytologic characteristics have not been well described. We report two cases of glycogen-rich clear cell carcinoma with corresponding fine needle aspiration(FNA) cytologic findings and compare them to infiltrating ductal carcinoma and other clear ceil malignancies with a review of literature. One was a 62-year-old woman exhibiting a palpable mass of the right breast. The smears showed atypical tight cell clusters and individually scattered single cells containing leanly or clear abundant cytoplasm with well defined cytoplasmic margins. Mild to moderate nuclear pleomorphism and a prominent nucleolus were present. The other was a 42-year-old woman who was admitted with a right breast mass. The smears showed moderately cellular, tightly cohesive tumor cells. The cytoplasmic outline was generally well demarcated. The tumor cells Contained foamy to clear abundant cytoplasm with large and small vacuoles. The nuclear pleomorphism was marked. Both tumors resected by modified radical mastectomy, were diagnosed as glycogen-rich clear cell carcinoma. Histologically, the clear cell nature of tumor cells were not characteristic enough to predict this type of the tumor. Some cytologic features can be distinguished other clear cell breast cancer from glycogen-rich carcinoma. Recognition of these unusual patterns in a breast FNAC should raise the suspicion of a clear cell carcinoma including glycogen-rich subtype. Cytological localization of glycogen using PAS and D-PAS staining may permit the correct Identification and differential diagnosis of this tumor.

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Expression of EMSY, a Novel BRCA2-link Protein, is Associated with Lymph Node Metastasis and Increased Tumor Size in Breast Carcinomas

  • Madjd, Zahra;Akbari, Mohammad Esmaeil;Zarnani, Amir Hassan;Khayamzadeh, Maryam;Kalantari, Elham;Mojtabavi, Nazanin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1783-1789
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    • 2014
  • Background: The EMSY gene encodes a BRCA2-binding partner protein that represses the DNA repair function of BRCA2 in non-hereditary breast cancer. Although amplification of EMSY gene has been proposed to have prognostic value in breast cancer, no data have been available concerning EMSY tissue expression patterns and its associations with clinicopathological features. Materials and Methods: In the current study, we examined the expression and localization pattern of EMSY protein by immunohistochemistry and assessed its prognostic value in a well-characterized series of 116 unselected breast carcinomas with a mean follow up of 47 months using tissue microarray technique. Results: Immunohistochemical expression of EMSY protein was detected in 76% of primary breast tumors, localized in nuclear (18%), cytoplasmic (35%) or both cytoplasmic and nuclear sites (23%). Univariate analysis revealed a significant positive association between EMSY expression and lymph node metastasis (p value=0.045) and larger tumor size (p value=0.027), as well as a non-significant relation with increased risk of recurrence (p value=0.088), whereas no association with patients' survival (log rank test, p value=0.482), tumor grade or type was observed. Conclusions: Herein, we demonstrated for the first time the immunostaining pattern of EMSY protein in breast tumors. Our data imply that EMSY protein may have impact on clinicipathological parameters and could be considered as a potential target for breast cancer treatment.