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Hepatitis C Virus NS4B

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https://www.readbyqxmd.com/read/28743875/hepatitis-c-virus-exploits-death-receptor-6-mediated-signaling-pathway-to-facilitate-viral-propagation
#1
Trang T D Luong, Giao V Q Tran, Dong-Jo Shin, Yun-Sook Lim, Soon B Hwang
The life cycle of hepatitis C virus (HCV) is highly dependent on host proteins for virus propagation. By transcriptome sequencing analysis, we identified host genes that were highly differentially expressed in HCV-infected cells. Of these candidates, we selected Death receptor 6 (DR6) for further characterization. DR6 is an orphan member of the tumor necrosis factor receptor superfamily. In the present study, we demonstrated that both mRNA and protein levels of DR6 were increased in the context of HCV replication...
July 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28727784/an-ns5a-single-optimized-method-to-determine-genotype-subtype-and-resistance-profiles-of-hepatitis-c-strains
#2
Elisabeth Andre-Garnier, Bernard Besse, Audrey Rodallec, Olivier Ribeyrol, Virginie Ferre, Caroline Luco, Laura Le Guen, Nathalie Bourgeois, Jérôme Gournay, Eric Billaud, François Raffi, Marianne Coste-Burel, Berthe-Marie Imbert-Marcille
The objective was to develop a method of HCV genome sequencing that allowed simultaneous genotyping and NS5A inhibitor resistance profiling. In order to validate the use of a unique RT-PCR for genotypes 1-5, 142 plasma samples from patients infected with HCV were analysed. The NS4B-NS5A partial region was successfully amplified and sequenced in all samples. In parallel, partial NS3 sequences were analyzed obtained for genotyping. Phylogenetic analysis showed concordance of genotypes and subtypes with a bootstrap >95% for each type cluster...
2017: PloS One
https://www.readbyqxmd.com/read/28588082/ire1%C3%AE-promotes-viral-infection-by-conferring-resistance-to-apoptosis
#3
Susan L Fink, Teshika R Jayewickreme, Ryan D Molony, Takao Iwawaki, Charles S Landis, Brett D Lindenbach, Akiko Iwasaki
The unfolded protein response (UPR) is an ancient cellular pathway that detects and alleviates protein-folding stresses. The UPR components X-box binding protein 1 (XBP1) and inositol-requiring enzyme 1α (IRE1α) promote type I interferon (IFN) responses. We found that Xbp1-deficient mouse embryonic fibroblasts and macrophages had impaired antiviral resistance. However, this was not because of a defect in type I IFN responses but rather an inability of Xbp1-deficient cells to undergo viral-induced apoptosis...
June 6, 2017: Science Signaling
https://www.readbyqxmd.com/read/28551625/intrahepatic-hcv-rna-level-and-genotype-1-independently-associate-with-hepatic-reticulon-3-expression
#4
Chih-Lang Lin, Rong-Nan Chien, Kung-Hao Liang, Po-Yuan Ke, Ya-Hui Huang, Chau-Ting Yeh
Background /Aim: Reticulon 3 (RTN3), resides predominantly in the endoplasmic reticulum and has opposite regulatory effects on Hepatitis C virus (HCV) replication through interacting with NS5A and NS4B proteins. This study aimed to unravel the actual effect of RTN3 on HCV replication. MATERIALS AND METHODS: A total of 115 HCV-related hepatocellular carcinoma patients receiving hepatectomy was enrolled in this study. The hepatic HCV RNA and RTN3 protein levels in the non-cancerous liver tissues were examined for clinical analysis...
June 2017: Anticancer Research
https://www.readbyqxmd.com/read/28545480/effects-of-hepatitis-c-virus-core-protein-and-nonstructural-protein-4b-on-the-wnt-%C3%AE-catenin-pathway
#5
Xiao-Hua Jiang, Yu-Tao Xie, Ya-Ping Cai, Jing Ren, Tao Ma
BACKGROUND: Hepatitis C virus (HCV) core protein and nonstructural protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis. RESULTS: Compared with the empty vector control, HCV core protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40...
May 25, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/28431572/hepatitis-c-virus-ns4b-protein-induces-epithelial-mesenchymal-transition-by-upregulation-of-snail
#6
Bicheng Hu, Shenggao Xie, Yuqian Hu, Wen Chen, Xiaofan Chen, Yi Zheng, Xinxing Wu
BACKGROUND: Chronic hepatitis C virus (HCV) infection is an important cause of hepatocellular carcinoma (HCC). Epithelial to mesenchymal transition (EMT) is a key process associated with tumor metastasis and poor prognosis. HCV infection, HCV core and NS5A protein could induce EMT process, but the role of NS4B on EMT remains poorly understood. METHODS: We overexpressed HCV NS4B protein in HepG2 cells or Huh7.5.1 cells infected by HCVcc, the E-cadherin expression, N-cadherin expression and the EMT-associated transcriptional factor Snail were determined...
April 21, 2017: Virology Journal
https://www.readbyqxmd.com/read/28170421/comprehensive-mapping-of-antigen-specific-t-cell-responses-in-hepatitis-c-virus-infected-patients-with-or-without-spontaneous-viral-clearance
#7
Chao Zhang, Rui Hua, Yuanyuan Cui, Shasha Wang, Hongqing Yan, Dongmei Li, Yonghong Zhang, Zhengkun Tu, Pei Hao, Xinyue Chen, Jin Zhong, Junqi Niu, Xia Jin
Elucidating protective immunity against HCV is important for the development of a preventative vaccine. We hypothesize that spontaneous resolution of acute HCV infection offers clue to protective immune responses, and that DAA therapy affects the quality and quantity of HCV-specific T cell responses. To test these hypotheses, we performed T cell epitope mapping in 111 HCV-infected individuals including 61 chronically HCV-1b (CHC-1b) infected, 24 chronically HCV-2a (CHC-2a) infected and 26 spontaneously recovered (SPR) patients with 376 overlapping peptides covering the entire HCV polyprotein...
2017: PloS One
https://www.readbyqxmd.com/read/27932311/daclatasvir-prevents-hepatitis-c-virus-infectivity-by-blocking-transfer-of-the-viral-genome-to-assembly-sites
#8
Bertrand Boson, Solène Denolly, Fanny Turlure, Christophe Chamot, Marlène Dreux, François-Loïc Cosset
BACKGROUND & AIMS: Daclatasvir is a direct-acting antiviral agent and potent inhibitor of NS5A, which is involved in replication of the hepatitis C virus (HCV) genome, presumably via membranous web shaping, and assembly of new virions, likely via transfer of the HCV RNA genome to viral particle assembly sites. Daclatasvir inhibits the formation of new membranous web structures and, ultimately, of replication complex vesicles, but also inhibits an early assembly step. We investigated the relationship between daclatasvir-induced clustering of HCV proteins, intracellular localization of viral RNAs, and inhibition of viral particle assembly...
March 2017: Gastroenterology
https://www.readbyqxmd.com/read/27923769/recombination-in-hepatitis-c-virus-is-not-uncommon-among-people-who-inject-drugs-in-kolkata-india
#9
Debanjali Gupta, Kallol Saha, Aritra Biswas, Rushna Firdaus, Monika Ghosh, Provash Chandra Sadhukhan
Recombination in RNA virus is a rare event in the survival and evolution to evade host immune system. This is increasing within high risk group population (HRG) due to super infection that occurs by continuous sharing of common drug equipment by HCV infected or HIV-HCV co-infected recurrent drug users. Recombination causes impediment to vaccine development and therapeutic intervention as standard HCV treatment is still genotype specific. Blood samples of 194 people who inject drugs (PWID) were collected from an Opioid Substitution Therapy Centre in Kolkata, India...
December 5, 2016: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/27678340/production-of-infectious-hcv-genotype-1b-virus-in-cell-culture-using-a-novel-set-of-adaptive-mutations
#10
Ken-Ichi Mori, Akihiro Matsumoto, Noboru Maki, Yuki Ichikawa, Eiji Tanaka, Shintaro Yagi
BACKGROUND: Despite the high prevalence of genotype 1b hepatitis C virus (HCV) among patients, a cell culture system that permits entire viral life cycle of genotype 1b isolates is limited. To develop a cell-cultured hepatitis C virus (HCVcc) of genotype 1b, the proper combination of HCV genomic variants and host cells is essential. HCV genomes isolated from patients with distinctive symptoms may provide the variants required to establish an HCVcc of genotype 1b. RESULTS: We first established subgenomic replicons in Huh7 cells using HCV cDNAs isolated from two patients: one with fulminant hepatitis after liver transplantation (TPF1) and another with acute hepatitis and moderate symptoms (sAH)...
September 27, 2016: BMC Microbiology
https://www.readbyqxmd.com/read/27620477/ptc725-an-ns4b-targeting-compound-inhibits-a-hepatitis-c-virus-genotype-3-replicon-as-predicted-by-genome-sequence-analysis-and-determined-experimentally
#11
Jason D Graci, Stephen P Jung, John Pichardo, Frederick Lahser, Xiao Tong, Zhengxian Gu, Joseph M Colacino
PTC725 is a small molecule NS4B-targeting inhibitor of hepatitis C virus (HCV) genotype (gt) 1 RNA replication that lacks activity against HCV gt2. We analyzed the Los Alamos HCV sequence database to predict susceptible/resistant HCV gt's according to the prevalence of known resistance-conferring amino acids in the NS4B protein. Our analysis predicted that HCV gt3 would be highly susceptible to the activity of PTC725. Indeed, PTC725 was shown to be active against a gt3 subgenomic replicon with a 50% effective concentration of ∼5 nM...
December 2016: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/27503576/isolation-and-characterization-of-hepatitis-c-virus-resistant-to-a-novel-phenanthridinone-derivative
#12
Wataru Ito, Masaaki Toyama, Mika Okamoto, Masanori Ikeda, Koichi Watashi, Takaji Wakita, Yuichi Hashimoto, Masanori Baba
BACKGROUND: The novel phenanthridinone derivative HA-719 has recently been identified as a highly potent and selective inhibitor of hepatitis C virus replication. To elucidate its mechanism of inhibition, we have isolated and analyzed a clone of hepatitis C virus replicon cells resistant to HA-719. METHODS: To isolate HA-719-resistant replicon cells, Huh-7 cells containing subgenomic hepatitis C virus replicons (genotype 1b) with a luciferase reporter (LucNeo#2) were cultured in the presence of G418 and escalating concentrations of HA-719...
August 8, 2016: Antiviral Chemistry & Chemotherapy
https://www.readbyqxmd.com/read/27489281/human-choline-kinase-%C3%AE-promotes-hepatitis-c-virus-rna-replication-through-modulation-of-membranous-viral-replication-complex-formation
#13
Mun-Teng Wong, Steve S Chen
UNLABELLED: Hepatitis C virus (HCV) infection reorganizes cellular membranes to create an active viral replication site named the membranous web (MW). The role that human choline kinase-α (hCKα) plays in HCV replication remains elusive. Here, we first showed that hCKα activity, not the CDP-choline pathway, promoted viral RNA replication. Confocal microscopy and subcellular fractionation of HCV-infected cells revealed that a small fraction of hCKα colocalized with the viral replication complex (RC) on the endoplasmic reticulum (ER) and that HCV infection increased hCKα localization to the ER...
October 15, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27318115/heterogeneous-genomic-locations-within-ns3-ns4a-and-ns4b-identified-for-genotyping-and-subtyping-of-hepatitis-c-virus-a-simple-genome-analysis-approach
#14
Sajid Mansoor, Aneela Javed, Amjad Ali, Atika Mansoor
Hepatitis C virus (HCV) displays excessive genetic heterogeneity and exists in several genotypes and subtypes. Characterizing these genotypes and subtypes becomes extremely important for diagnostic and epidemiological reasons. Present study analyzed HCV genome using a simple genome analysis approach. We combined manual sectioning of a reference genome alignment (RA) followed by a comprehensive comparative phylogenetic analysis. The main aim was to identify heterogeneous locations on HCV genome suitable for genotyping/subtyping...
October 2016: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/27315218/hcv-ns4b-targets-scribble-for-proteasome-mediated-degradation-to-facilitate-cell-transformation
#15
Bo Hu, Shanshan Li, Zhanfeng Zhang, Shenggao Xie, Yuqian Hu, Xianzhang Huang, Yi Zheng
Hepatitis C virus (HCV) nonstructural protein 4B (NS4B) is a multi-transmembrane protein, but little is known about how NS4B contributes to HCV replication and tumorigenesis. Its C-terminal domain (CTD) has been shown to associate with intracellular membrane, and we have previously shown that NS4B CTD contains a class I PDZ-binding motif (PBM). Here, we demonstrated that NS4B PBM interacts with the PDZ-containing tumor suppressor protein, Scribble, using immunofluorescence and co-immunoprecipitation assays, and this interaction requires at least three contiguous PDZ domains of Scribble...
September 2016: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
https://www.readbyqxmd.com/read/27277863/genome-wide-analysis-for-identification-of-adaptive-diversification-between-hepatitis-c-virus-subtypes-1a-and-1b
#16
Yan Li, Ruirui Wang, Xiaogang Du, Mingwang Zhang, Meng Xie
Hepatitis C virus (HCV) is a major cause of liver disease and has been estimated to infect approximately 2%-3% of the world's population. HCV genotype 1 is the subject of intense research and clinical investigations because of its worldwide prevalence and poor access to treatment for patients in developing countries and marginalized populations. The predominant subtypes 1a and 1b of HCV genotype 1 present considerable differences in epidemiological features. However, the genetic signature underlying such phenotypic functional divergence is still an open question...
July 2016: Canadian Journal of Microbiology
https://www.readbyqxmd.com/read/27273268/a-highly-specific-and-sensitive-hepatitis-c-virus-antigen-enzyme-immunoassay-for-one-step-diagnosis-of-viremic-hepatitis-c-virus-infection
#17
Ke-Qin Hu, Wei Cui
UNLABELLED: The current standard in diagnosing hepatitis C virus (HCV) infection requires two sequential steps: anti-HCV test to screen, followed by HCV RNA reverse-transcription polymerase chain reaction to confirm viremic HCV (V-HCV) infection. HCV core antigen tests provided potential for possible one-step diagnosis. However, low sensitivity and specificity limit their clinical utility. The present study developed a novel HCV antigens enzyme immunoassay (HCV-Ags EIA) and assessed its sensitivity, specificity, and utility for one-step diagnosis of V-HCV infection using 365 serum specimens, including 176 without and 189 with V-HCV infection...
August 2016: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
https://www.readbyqxmd.com/read/27252526/interaction-between-nonstructural-proteins-ns4b-and-ns5a-is-essential-for-proper-ns5a-localization-and-hepatitis-c-virus-rna-replication
#18
Avik Biswas, Jason Treadaway, Timothy L Tellinghuisen
UNLABELLED: The hepatitis C virus NS5A protein is tethered to cellular membranes via an amphipathic amino-terminal helix that is inserted in-plane into the outer endoplasmic reticulum (ER)-derived membrane leaflet. The charged face of the helix faces the cytoplasm and may contribute to interactions involved in replicase assembly and function. Using an aggressive charge flip mutagenesis strategy, we identified a number of essential residues for replication on the charged face of the NS5A anchor and identified a double charge face mutant that is lethal for RNA replication but generates suppressor mutations in the carboxy-terminal helix of the NS4B protein...
August 15, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27240954/inhibition-of-hcv-replication-by-humanized-single-domain-transbodies-to-ns4b
#19
Kittirat Glab-Ampai, Aijaz Ahmad Malik, Monrat Chulanetra, Jeeraphong Thanongsaksrikul, Kanyarat Thueng-In, Potjanee Srimanote, Pongsri Tongtawe, Wanpen Chaicumpa
NS4B of hepatitis C virus (HCV) initiates membrane web formation, binds RNA and other HCV proteins for viral replication complex (RC) formation, hydrolyses NTP, and inhibits innate anti-viral immunity. Thus, NS4B is an attractive target of a novel anti-HCV agent. In this study, humanized-nanobodies (VHs/VHHs) that bound to recombinant NS4B were produced by means of phage display technology. The nanobodies were linked molecularly to a cell penetrating peptide, penetratin (PEN), for making them cell penetrable (become transbodies)...
August 5, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27233794/cell-free-expression-purification-and-membrane-reconstitution-for-nmr-studies-of-the-nonstructural-protein-4b-from-hepatitis-c-virus
#20
Marie-Laure Fogeron, Vlastimil Jirasko, Susanne Penzel, David Paul, Roland Montserret, Clément Danis, Denis Lacabanne, Aurélie Badillo, Jérôme Gouttenoire, Darius Moradpour, Ralf Bartenschlager, François Penin, Beat H Meier, Anja Böckmann
We describe the expression of the hepatitis C virus nonstructural protein 4B (NS4B), which is an integral membrane protein, in a wheat germ cell-free system, the subsequent purification and characterization of NS4B and its insertion into proteoliposomes in amounts sufficient for multidimensional solid-state NMR spectroscopy. First spectra of the isotopically [(2)H,(13)C,(15)N]-labeled protein are shown to yield narrow (13)C resonance lines and a proper, predominantly α-helical fold. Clean residue-selective leucine, isoleucine and threonine-labeling is demonstrated...
June 2016: Journal of Biomolecular NMR
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