• Title/Summary/Keyword: nucleoprotein

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Role of Chromatin Structure in HMRE Mediated Transcriptional Repression of the HSP82 Heat Shock Gene

  • Lee, See-Woo;Gross, David S.
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.40-48
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    • 1996
  • We have examined the chromatin structure of the HMRE/HSP82 and HMRa/HSP82 allels using three complementary approaches : DNase I chromating footprinting, micrococcal nuclease (MNase) nucleosome-protected ladder assay, and an in vivo E. coli dam methylase accessibility assay. The footprinting results indicate that the promoter and silencer sequences are assembled into nucleoprotein complexes which exhibit no detectable change in structure, despite a 70-fold range in expression levels. In addition, the promoter region of the HMRa/HSP82 allele is cleaved randomly by MNase in all cases, indicating the absence of anonical nucleosomes over this region irrespective of SIR4 or heat-shock. Finally, no discernible difference in the accessibility of the HMRE/HSP82 locus to dam methylase in SIR4 vs. sir4 cells was seenm which again suggests that the chromatin structure of HMRE/HSP82 allele is identical regardless of SIR4. Altogether, our results indicate that in contrast to other observations of the silent mating-type loci, no discernible structural alteration is detected at either HMR/HSP82 allele regardless of SIR genetic background or transcriptional state of the gene.

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Foamy Virus Integrase in Development of Viral Vector for Gene Therapy

  • Kim, Jinsun;Lee, Ga-Eun;Shin, Cha-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1273-1281
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    • 2020
  • Due to the broad host suitability of viral vectors and their high gene delivery capacity, many researchers are focusing on viral vector-mediated gene therapy. Among the retroviruses, foamy viruses have been considered potential gene therapy vectors because of their non-pathogenicity. To date, the prototype foamy virus is the only retrovirus that has a high-resolution structure of intasomes, nucleoprotein complexes formed by integrase, and viral DNA. The integration of viral DNA into the host chromosome is an essential step for viral vector development. This process is mediated by virally encoded integrase, which catalyzes unique chemical reactions. Additionally, recent studies on foamy virus integrase elucidated the catalytic functions of its three distinct domains and their effect on viral pathogenicity. This review focuses on recent advancements in biochemical, structural, and functional studies of foamy virus integrase for gene therapy vector research.

Generation of heterologous proteins-expressing recombinant snakehead rhabdoviruses (rSHRVs) using reverse genetics

  • Kwak, Jun Soung;Ryu, Sujeong;Kim, Ki Hong
    • Journal of fish pathology
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    • v.33 no.2
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    • pp.163-169
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    • 2020
  • Snakehead rhabdovirus (SHRV) is different from other fish novirhabdoviruses such as viral hemorrhagic septicemia virus (VHSV), infectious hematopoietic necrosis virus (IHNV), and hirame rhabdovirus (HIRRV) in that it replicates at high temperatures. Therefore, the delivery of foreign proteins to fish living at high water temperature would be possible by using recombinant SHRVs. In the present study, to evaluate the possible use of SHRV as a vehicle for foreign proteins delivery, we generated a recombinant SHRV that contains an enhanced-GFP (eGFP) gene between nucleoprotein (N) and phosphoprotein (P) genes (rSHRV-A-eGFP), and another recombinant SHRV expressing two heterologous genes by inserting an eGFP gene between N and P genes, and mCherry gene between P and M genes (rSHRV-AeGFP-BmCherry). Epithelioma papulosum cyprini (EPC) cells infected with the recombinant SHRVs showed strong fluorescence(s), suggesting the possible availability of recombinant SHRVs for the development of combined vaccines by expressing multiple foreign antigens.

Comparative analysis of antibody responses to BNT162b2, ChAdOx1, and CoronaVac vaccines in the Albanian population over the pandemic years 2021 to 2022

  • Genc Sulcebe;Margarita Kurti-Prifti;Erkena Shyti;Jonida Dashi-Pasholli;Fabian Cenko;Alban Ylli
    • Clinical and Experimental Vaccine Research
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    • v.13 no.1
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    • pp.63-67
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    • 2024
  • This repeated cross-sectional study with two independent sample populations compared the antibody response to severe acute respiratory syndrome coronavirus 2 vaccines in Albania in July-August 2021 and 2022. In 2021, it found higher anti-spike-1 seropositivity and antibody levels in fully vaccinated individuals, especially with BNT162b2 and ChAdOx1 and to a lesser degree with CoronaVac. By 2022, all single-dose recipients showed high antibody responses, suggesting natural infection-enhanced immunity. The study indicates a significant evolution in the antibody response to different coronavirus disease 2019 vaccines and suggests that a single vaccine dose, coupled with natural infection, might suffice to maintain adequate immunity levels in an endemic scenario.

A New Rhabdovirus (HRV-like) Isolated in Korea from Cultured Japanese Flounder Paralichthys olivaceus (양식산 넙치로부터 HRV-like Rhabdovirus의 분리)

  • Oh, Myung-Joo;Choi, Tae-Jin
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.129-136
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    • 1998
  • In March 1997, a new rhabdovirus was isolated from moribund cultured Japanese flounder Paralichthys olivaceus in sea water tank and cage culture systems in Kyung-Nam and Chun-Nam province, Korea. At temperature $15^{\circ}C$ the virus replicated and induced cytopathic effects (CPE), which progressed to eventual cytolysis, in susceptible cell lines, including RTG-2 and EPC. The CHES-214 cell line was refractory. Virus particles were bullet-shaped and measured $70nm{\times}100$ to 150 nm in size. The isolate was sensitive to pH 3, to diethyl ether, and to heat ($50^{\circ}C$ 5 min, $60^{\circ}C$ 1 min). Viral replication was not inhibited by $10^{-4}$ M 5-iododeoxyuridine. Virus infectivity was reduced by anti-HRV (8401-H) rabbit serum, but can not reduced by antisera against infectious hematopoietic necrosis virus (IHNV), chum salmon reovirus (CSV), retrovirus of salmonid (RVS) and infectious pancreatic necrosis virus (IPNV). HRV virus antigen was detected by fluorescent antibody test (FAT) in the cytoplasm of infected EPC cell. Purified isolates virions were composed of: polymerase (L), glycoprotein (G), nucleoprotein (N) and 2 matrix proteins (M1 and M2). Based upon their relative mobilities, the estimated molecular weights of the proteins were: L, 160 kDa; G, 55 kDa; N, 45 kDa; M1, 26 kDa; and M2, 22 kDa.

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Molecular Characterization of an Avian-origin Reassortant H7N1 Influenza Virus (조류 유래 재조합 H7N1 인플루엔자 바이러스의 분자적 특성 규명)

  • Sun-Woo Yoon
    • Journal of Life Science
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    • v.33 no.8
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    • pp.605-611
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    • 2023
  • Recently, sporadic cases of human infection by genetic reassortants of H7Nx influenza A viruses have been reported; such viruses have also been continuously isolated from avian species. In this study, A/wild bird/South Korea/sw-anu/2023, a novel reassortant of the H7N1 avian influenza virus, was analyzed using full-genome sequencing and molecular characterization. Phylogenetic analysis showed that A/wild bird/South Korea/sw-anu/2023 belonged to the Eurasian lineage of H7Nx viruses. The polymerase basic (PB)2, PB1, polymerase acidic (PA), and nucleoprotein (NP) genes of these viruses were found to be closely related to those of avian influenza viruses isolated from wild birds, while the hemagglutinin (HA), neuraminidase (NA), matrix (M), and nonstructural (NS) genes were similar to those of avian influenza viruses isolated from domestic ducks. In addition, A/wild bird/South Korea/sw-anu/2023 also had a high binding preference for avian-specific glycans in the solid-phase direct binding assay. These results suggest the presence of a new generation of H7N1 avian influenza viruses in wild birds and highlight the reassortment of avian influenza viruses found along the East Asian-Australasian flyway. Overall, H7Nx viruses circulate worldwide, and mutated H7N1 avian viruses may infect humans, which emphasizes the requirement for continued surveillance of the H7N1 avian influenza virus in wild birds and poultry.

Production of monoclonal antibodies against viral hemorrhagic septicemia virus (VHSV, genotype IVa) from olive flounder (넙치 유래 viral hemorrhagic septicemia virus (VHSV, genotype IVa)에 대한 단클론 항체 생산)

  • Jeong, Ha-Na;Jang, Min-Seok;Oh, Myung-Joo;Kim, Wi-Sik
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.149-154
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    • 2017
  • Mouse monoclonal antibodies (MAbs) were produced by using viral hemorrhagic septicemia virus (VHSV, genotype IVa) as an immunogen, isolated from diseased olive flounder (Paralichthys olivaceus). Four hybridoma clones secreting MAbs against VHSV were established. The MAbs were recognized the nucleoprotein (MAb 4), phosphoprotein (MAb 1) and matrix protein (MAbs 2 and 3) of VHSV by western blot analysis. Among them, the MAbs 1 and 4 strongly reacted with the VHSV-infected FHM cells, but not normal FHM cells. In enzyme linked immunosorbent assay, the four MAbs reacted with the VHSV, but not different six fish viruses (infectious hematopoietic necrosis virus, hirame rhabdovirus, spring viraemia of carp virus, infectious pancreatic necrosis virus, marine birnavirus and nervous necrosis virus). These results indicate that the MAbs are useful for diagnosis of VHSV infection.

PB-Overexpression of OsZn15, a CCCH-tandem zinc finger protein, increases drought tolerance in rice

  • Seong, So Yoon;Jung, Harin;Choi, Yang Do;Kim, Ju-Kon
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.115-115
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    • 2017
  • Zinc finger proteins constitute a large family which has been studied to have various functions in different organisms. Tandem CCCH zinc finger proteins (TZFs), members of the zinc finger protein family, are known to participate as post-transcriptional regulators of gene expression in eukaryotes. Here, we showed that the OsZn15, a gene for tandem CCCH zinc finger protein, is induced by abiotic stress and its overexpression in transgenic rice plants (PGD1:OsZn15) gains higher drought tolerance. Gene expression analysis of promoter:GFP plants revealed that OsZn15 is specifically expressed in anther and embryo, but not in vegetative organs. In-field evaluation, grain yield was higher in the PGD1:OsZn15 than nontransgenic plants under drought conditions. Interestingly, OsZn15 is shown to not only localize at nucleus but also co-localize with both processing bodies (PB) and stress granules (SG), two messenger ribo-nucleoprotein complexes which are known to activate by forming cytoplasmic foci under stress conditions. In sum, these results suggest that OsZn15 increases drought stress tolerance of rice probably by participating in RNA turnover in PB and SG.

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Partial Biological and Molecular Characterization of Tomato yellow fruit ring virus Isolates from Potato

  • Pourrahim, Reza;Golnaraghi, Alireza;Farzadfar, Shirin;Ohshima, Kazusato
    • The Plant Pathology Journal
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    • v.28 no.4
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    • pp.390-400
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    • 2012
  • Eight potato-producing provinces of Iran were surveyed during the growing seasons of 2004-2006 to detect the presence of Tomato yellow fruit ring virus (TYFRV), a tentative species in the genus Tospovirus. A total of 1,957 potato leaf samples were collected from plants with tospovirus-like symptoms of chlorotic or necrotic spots, chlorosis and necrosis. The samples were tested by enzyme-linked immunosorbent assay using TYFRV-specific antibodies. Among those tested, 498 samples (25.4%) were found to be infected with the virus. The virus was detected in 72.4% of the potato fields in all provinces surveyed. Thirteen potato isolates of TYFRV were selected for further biological and molecular studies. Based on their reactions on Nicotiana tabacum plants, the isolates were separated into two groups, namely L (local infection) and N (systemic infection). The nucleotide sequences of the nucleoprotein (N) genes of the isolates were determined and compared with the homologous sequences in Genbank. No recombination evidence was found in the isolates using different recombination-detecting programs. In the phylogenetic tree, the potato isolates fell into two major groups: IRN-1 and IRN-2 corresponding to the two biologically separated groups. This study shows for the first time the biological and phylogenetic relationships of geographically distant TYFRV isolates from potatoes in the mid-Eurasian country of Iran.

Constructing Proteome Reference Map of the Porcine Jejunal Cell Line (IPEC-J2) by Label-Free Mass Spectrometry

  • Kim, Sang Hoon;Pajarillo, Edward Alain B.;Balolong, Marilen P.;Lee, Ji Yoon;Kang, Dae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1124-1131
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    • 2016
  • In this study, the global proteome of the IPEC-J2 cell line was evaluated using ultra-high performance liquid chromatography coupled to a quadrupole Q Exactive Orbitrap mass spectrometer. Proteins were isolated from highly confluent IPEC-J2 cells in biological replicates and analyzed by label-free mass spectrometry prior to matching against a porcine genomic dataset. The results identified 1,517 proteins, accounting for 7.35% of all genes in the porcine genome. The highly abundant proteins detected, such as actin, annexin A2, and AHNAK nucleoprotein, are involved in structural integrity, signaling mechanisms, and cellular homeostasis. The high abundance of heat shock proteins indicated their significance in cellular defenses, barrier function, and gut homeostasis. Pathway analysis and annotation using the Kyoto Encyclopedia of Genes and Genomes database resulted in a putative protein network map of the regulation of immunological responses and structural integrity in the cell line. The comprehensive proteome analysis of IPEC-J2 cells provides fundamental insights into overall protein expression and pathway dynamics that might be useful in cell adhesion studies and immunological applications.