• Title/Summary/Keyword: viral protein

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Pathway Analysis in HEK 293T Cells Overexpressing HIV-1 Tat and Nucleocapsid

  • Lee, Min-Joo;Park, Jong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1103-1108
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    • 2009
  • The human immunodeficiency virus (HIV)-l protein Tat acts as a transcription transactivator that stimulates expression of the infected viral genome. It is released from infected cells and can similarly affect neighboring cells. The nucleocapsid is an important protein that has a related significant role in early mRNA expression, and which contributes to the rapid viral replication that occurs during HIV-1 infection. To investigate the interaction between the Tat and nucleocapsid proteins, we utilized cDNA micro arrays using pTat and flag NC cotransfection in HEK 293T cells and reverse transcription-polymerase chain reaction to validate the micro array data. Four upregulated genes and nine downregulated genes were selected as candidate genes. Gene ontology analysis was conducted to define the biological process of the input genes. A proteomic approach using PathwayStudio determined the relationship between Tat and nucleocapsid; two automatically built pathways represented the interactions between the upregulated and downregulated genes. The results indicate that the up- and downregulated genes regulate HIV-1 replication and proliferation, and viral entry.

Host Cell-Intrinsic Antiviral Defense Induced by Type I Interferons

  • Asano, Atsushi
    • Korean Journal of Agricultural Science
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    • v.35 no.2
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    • pp.177-182
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    • 2008
  • Type I Interferons (IFNs) are potent antiviral cytokines that modulate both innate immunity and adaptive immunity. Type I IFNs are immediately induced by viral infection, and stimulate production of a broad range of gene products such as double-stranded RNA-activated protein kinase (PKR), 2' 5'-oligoadenylate synthetase (OAS)/RNaseL and Mx GTPases. These proteins inhibit viral replication in host cells. Type I IFNs, in turn, lead to antiviral state at early phase of viral infection. We provide an overview of the knowledge of IFN-inducible antiviral proteins conserved in vertebrates.

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Smad4 Expression in Hepatocellular Carcinoma Differs by Hepatitis Status

  • Yao, Lei;Li, Fu-Jun;Tang, Zhi-Qiang;Gao, Shuang;Wu, Qe-Quan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1297-1303
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    • 2012
  • Aims: Primary hepatocellular carcinoma (HCC) is a common malignancy often related to hepatitis viral infection. Smad4 is known to mediate the TGF-${\beta}$ pathway to suppress tumorigenesis. However, the function of Smad4 in HCC is still controversial. In this study we compared levels of Smad4 in HCC tissues with or without hepatitis virus infection and adjacent normal-appearing liver. Methods: Samples from HCC patients were analyzed for Smad4 protein and mRNA expression by immunohistochemistry (IHC), RT-PCR and Western blotting. Results: We found that tumor tissues expressed less Smad4 mRNA and protein than the adjacent tissues. Most HCC tumor tissues were negative for Smad4 in IHC staining, while the majority of adjacent tissues were positively stained. Interestingly, protein levels were higher in HCC tissues with viral hepatitis than those without virus infection. Suppression of expression appeared closely related to HCC, so that Smad4 appears to function as a tumor suppressor gene (TSG). Conclusion: Patients with hepatitis viral infection, at higher risk for HCC, exhibited increased Smad4 protein expression suggesting hepatitis virus may modulate Smad4 expression, which is functionally distinct from its putative role as a TSG. Smad4 expression may thus be an applicable marker for diagnosis and/or a target to develop therapeutic agents for HCC.

Druggability for COVID-19: in silico discovery of potential drug compounds against nucleocapsid (N) protein of SARS-CoV-2

  • Ray, Manisha;Sarkar, Saurav;Rath, Surya Narayan
    • Genomics & Informatics
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    • v.18 no.4
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    • pp.43.1-43.13
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    • 2020
  • The coronavirus disease 2019 is a contagious disease and had caused havoc throughout the world by creating widespread mortality and morbidity. The unavailability of vaccines and proper antiviral drugs encourages the researchers to identify potential antiviral drugs to be used against the virus. The presence of RNA binding domain in the nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) could be a potential drug target, which serves multiple critical functions during the viral life cycle, especially the viral replication. Since vaccine development might take some time, the identification of a drug compound targeting viral replication might offer a solution for treatment. The study analyzed the phylogenetic relationship of N protein sequence divergence with other 49 coronavirus species and also identified the conserved regions according to protein families through conserved domain search. Good structural binding affinities of a few natural and/or synthetic phytocompounds or drugs against N protein were determined using the molecular docking approaches. The analyzed compounds presented the higher numbers of hydrogen bonds of selected chemicals supporting the drug-ability of these compounds. Among them, the established antiviral drug glycyrrhizic acid and the phytochemical theaflavin can be considered as possible drug compounds against target N protein of SARS-CoV-2 as they showed lower binding affinities. The findings of this study might lead to the development of a drug for the SARS-CoV-2 mediated disease and offer solution to treatment of SARS-CoV-2 infection.

PVY Resistant Transgenic Potato Plants (cv Claustar) Expressing the Viral Coat Protein

  • Gargouri-Bouzid Radhia;Jaoua Leila;Mansour Riadh Ben;Hathat Yemna;Ayadi Malika;Ellouz Radhouane
    • Journal of Plant Biotechnology
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    • v.7 no.3
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    • pp.143-148
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    • 2005
  • The coat protein mediated resistance to potato virus Y is assessed here in transgenic potato plants (Solanum tuberosum L., cv Claustar). Therefore, the corresponding cDNA from tunisian isolate of the virus was cloned into Agrobacterium tumefaciens binary vector. The transgenic lines were subsequently analysed for the presence and expression of the transgene. The CP cDNA copy number was determined for kanamycin resistant plants. Three selected transgenic lines and their S1 progeny resulting from tuber germination showed a high protection level against the virus. These data appear to support the hypothesis that the virus resistance is mediated by the translated viral coat protein expressed in transgenic potato lines.

Complementary DNA Cloning and Nucleotide Sequence Analysis of Coat Protein Gene from TMV Tomato Strain (토마토에서 분리된 담배 모자이크 바이러스 외피단백질 유전자의 cDNA 클로닝 및 염기서열 분석)

  • 이청호;이영기;강신웅;박은경
    • Journal of the Korean Society of Tobacco Science
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    • v.18 no.2
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    • pp.101-106
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    • 1996
  • Tobacco mosaic virus (TMV) tomato strain was isolated from tomato "Seo-Kwang" in Korea. The virion was purified by density gradient centrifugation, and total viral RNA was isolated from the purified particles. Coat protein (CP) cDNA of the virus was synthesized by RT-PCR, and the purified cDNA fragment was subcloned to pBluescript II SK-. The analysis of nucleotide sequence showed that this cDNA was 693 nucleotides long from the insert of clone p1571 and p1572 which contain complete codons of the viral coat protein gene (474 nucleotides) and 3' untranslated region. The nucleotides of coat protein encoding cDNA of the strain were 6 nucleotides less than that of TMV common strain isolated from tobacco plant in Korea. The CP gene showed 70% maximum homology with that of the common strain in the nucleotide level and 86% maximum homology in amino acid level.cid level.

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Recent Advances and Future Strategy in Gene Delivery System (유전자 수송계의 현재까지의 연구동향 및 앞으로의 개발전략)

  • Choi, Woo-Jeong;Kim, Chong-Kook
    • Journal of Pharmaceutical Investigation
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    • v.30 no.1
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    • pp.1-12
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    • 2000
  • Gene therapy is a method for the treatment of diseases with introducing the gene-engineered materials into a patient with gene-deficiency disease (e.g. cystic fibrosis) or cancer to produce a therapeutic protein in a patient's cells. Successful gene therapy requires establishing both gene expression systems and delivery systems. Viral and non-viral vectors have been used for gene delivery. Viral vectors have a high transfection efficiency, but are limited in relations to issues of safety, toxicity and immunogenecity. Non-viral vectors are easy to prepare and relatively safe. However, non-viral vectors have a low transfection efficiency. Cationic liposomes are the most available among non-viral vectors. Cationic liposomes have been used to transfect cells both in vitro and in vivo experiments. Besides, several formulations containing cationic lipid are being used in clinical trials in cases of cystic fibrosis or cancer. A crucial subject to the further development of gene delivery vectors will be a long-term gene expression with following characteristics; protecting and deliverying DNA efficiently, non-toxic and non-immunogenic, and easy to produce in large scale.

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The Role of Noncoding Region in Hantaan Viral S Genome for Expression of Nucleocapsid Protein (한탄바이러스 Nucleocapsid Protein 발현에 있어 S Genome 내 Noncoding Region의 역할)

  • Yu, Cheong-Hee;Lee, Yeon-Seung;Lee, Ho-Dong;Park, Chan;Park, Keun-Yong;Lee, Pyung-Woo
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.39-49
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    • 2000
  • The genome of Hantaan virus, the prototype of the hantavirus genus, is composed of three segmented, single stranded negative sense RNA genome. The 5' and 3' termini of the Hantaan virus RNA genome contain noncoding regions (NCRs) that are highly conserved and complementary to form panhandle structures. There are some reports that these NCRs seems to control gene expression and viral replication in influenza virus and vesicular stomatitis virus. In this study, we examined whether NCRs in Hantaan virus playa role in expression of the viral nucleocapsid protein (Np) and foreign (luciferase) gene. The 5' and/or 3' NCR-deleted mutants were constructed and analysed. The Np expression of 5' NCR-deleted clone was similar to that of the clone containing full S genome. In the case of 3' NCR-deleted clone, it showed 40% reduction. To investigate the role of NCR in foreign gene expression, the clones which are replaced ORF of Hantaan viral Np gene with that of luciferase gene were constructed. The results were similar to those of the experiments using Np gene. These results suggest that 3' NCR is more important than 5' NCR in protein expression. To find out a critical region of 3' NCR in protein expression, several clones with a deleted part of 3' NCR were constructed and analyzed. The deletion of the conserved region in 3' NCR showed $20{\sim}30%$ decrease in Np expression. However there were no change in luciferase activities between clones with or without non-conserved region of 3' NCR. These results suggest that the 3' NCR of Hantaan virus S genome, especially conserved region in 3' NCR, plays an important role in the expression of Hantaan viral Np and foreign genes.

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Identification of a Cellular Protein Interacting with RNA Polymerase of Hepatitis C Virus

  • Park, Kyu-Jin;Choi, Soo-Ho;Koh, Moon-Soo;Kim, Sung-Wan;Hwang, Soon-Bong
    • BMB Reports
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    • v.33 no.1
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    • pp.59-62
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    • 2000
  • Hepatitis C virus (HCV) nonstructural 5B (NS5B) protein is an RNA-dependent RNA polymerase (RdRp). To determine whether it can contribute to viral replication by interaction with cellular proteins, the yeast two-hybrid screening system was employed to screen a human liver cDNA library. Using the HCV NS5B as a bait, we have isolated positive clones encoding a cellular protein. The NS5B interacting protein, 5BIP, is a novel cellular protein of 170 amino acids. Interaction of the HCV NS5B protein with 5BIP was confirmed by a protein-protein blotting assay. Recently, we have demonstrated that NS5B possesses an RdRp activity and thus it is possible that 5BIP, in association with NS5B, plays a role in HCV replication.

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