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Flavivirus NS2A

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https://www.readbyqxmd.com/read/27746901/illustrating-and-homology-modeling-the-proteins-of-the-zika-virus
#1
Sean Ekins, John Liebler, Bruno J Neves, Warren G Lewis, Megan Coffee, Rachelle Bienstock, Christopher Southan, Carolina H Andrade
The Zika virus (ZIKV) is a flavivirus of the family Flaviviridae, which is similar to dengue virus, yellow fever and West Nile virus. Recent outbreaks in South America, Latin America, the Caribbean and in particular Brazil have led to concern for the spread of the disease and potential to cause Guillain-Barré syndrome and microcephaly. Although ZIKV has been known of for over 60 years there is very little in the way of knowledge of the virus with few publications and no crystal structures. No antivirals have been tested against it either in vitro or in vivo...
2016: F1000Research
https://www.readbyqxmd.com/read/27665292/a-simple-method-for-developing-an-infectious-cdna-clone-of-japanese-encephalitis-virus
#2
Hao Zheng, Xuchen Zheng, Wu Tong, Fei Liu, Chao Liang, Tao Wang, Fei Gao, Liwei Li, Tongling Shan, Guoxin Li, Guangzhi Tong
Flavivirus cDNA clones frequently demonstrate genetic instability in transformed bacteria, which hampers the construction and manipulation of cDNAs for infectious flaviviruses. In this study, we developed a stable, full-length cDNA clone, pJEHEN, of a GI JEV strain HEN0701 using a medium-copy-number pBR322 vector and propagating cDNA clones at room temperature. The virus vJEHEN recovered from the infectious clone was indistinguishable from the parent virus HEN0701 with respect to plaque morphology, growth kinetics, and virulence characteristics...
September 24, 2016: Virus Genes
https://www.readbyqxmd.com/read/27296393/predicting-zika-virus-structural-biology-challenges-and-opportunities-for-intervention
#3
Bryan D Cox, Richard A Stanton, Raymond F Schinazi
BACKGROUND: Zika virus is an emerging crisis as infection is implicated in severe neurological disorders-Guillain-Barré syndrome and fetal microcephaly. There are currently no treatment options available for Zika virus infection. This virus is part of the flavivirus genus and closely related to Dengue Fever Virus, West Nile Virus, and Japanese Encephalitis Virus. Like other flaviviruses, the Zika virus genome encodes three structural proteins (capsid, precursor membrane, and envelope) and seven nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5)...
August 2015: Antiviral Chemistry & Chemotherapy
https://www.readbyqxmd.com/read/27169727/population-genomics-of-dengue-virus-serotype-4-insights-into-genetic-structure-and-evolution
#4
Vaishali P Waman, Sunitha Manjari Kasibhatla, Mohan M Kale, Urmila Kulkarni-Kale
The spread of dengue disease has become a global public health concern. Dengue is caused by dengue virus, which is a mosquito-borne arbovirus of the genus Flavivirus, family Flaviviridae. There are four dengue virus serotypes (1-4), each of which is known to trigger mild to severe disease. Dengue virus serotype 4 (DENV-4) has four genotypes and is increasingly being reported to be re-emerging in various parts of the world. Therefore, the population structure and factors shaping the evolution of DENV-4 strains across the world were studied using genome-based population genetic, phylogenetic and selection pressure analysis methods...
August 2016: Archives of Virology
https://www.readbyqxmd.com/read/27053551/transmembrane-domains-of-ns2b-contribute-to-both-viral-rna-replication-and-particle-formation-in-japanese-encephalitis-virus
#5
Xiao-Dan Li, Cheng-Lin Deng, Han-Qing Ye, Hong-Lei Zhang, Qiu-Yan Zhang, Dong-Dong Chen, Pan-Tao Zhang, Pei-Yong Shi, Zhi-Ming Yuan, Bo Zhang
UNLABELLED: Flavivirus nonstructural protein 2B (NS2B) is a transmembrane protein that functions as a cofactor for viral NS3 protease. The cytoplasmic region (amino acids 51 to 95) alone of NS2B is sufficient for NS3 protease activity, whereas the role of transmembrane domains (TMDs) remains obscure. Here, we demonstrate for the first time that flavivirus NS2B plays a critical role in virion assembly. Using Japanese encephalitis virus (JEV) as a model, we performed a systematic mutagenesis at the flavivirus conserved residues within the TMDs of NS2B...
June 15, 2016: Journal of Virology
https://www.readbyqxmd.com/read/26377679/proteolytic-cleavage-analysis-at-the-murray-valley-encephalitis-virus-ns1-2a-junction
#6
COMPARATIVE STUDY
Siti Nor Khadijah Addis, Eva Lee, Jayaram Bettadapura, Mario Lobigs
BACKGROUND: Our understanding of the proteolytic processing events at the NS1-2A junction in the flavivirus polyprotein has not markedly progressed since the early work conducted on dengue virus (DENV). This work identified an octapeptide sequence located immediately upstream of the cleavage site thought to be important in substrate recognition by an as yet unknown, endoplasmic reticulum-resident host protease. Of the eight amino acid recognition sequence, the highly conserved residues at positions P1, P3, P5, P7 and P8 (with respect to N-terminus of NS2A) are particularly sensitive to amino acid substitutions in terms of DENV NS1-NS2A cleavage efficiency; however, the role of the octapeptide in efficient NS1 and NS2A production of other flaviviruses has not been experimentally addressed...
2015: Virology Journal
https://www.readbyqxmd.com/read/25915765/profiling-of-viral-proteins-expressed-from-the-genomic-rna-of-japanese-encephalitis-virus-using-a-panel-of-15-region-specific-polyclonal-rabbit-antisera-implications-for-viral-gene-expression
#7
Jin-Kyoung Kim, Jeong-Min Kim, Byung-Hak Song, Sang-Im Yun, Gil-Nam Yun, Sung-June Byun, Young-Min Lee
Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is closely related to West Nile (WN), yellow fever (YF), and dengue (DEN) viruses. Its plus-strand genomic RNA carries a single open reading frame encoding a polyprotein that is cleaved into three structural (C, prM/M, and E) and at least seven nonstructural (NS1/NS1', NS2A, NS2B, NS3, NS4A, NS4B, and NS5) proteins, based on previous work with WNV, YFV, and DENV. Here, we aimed to profile experimentally all the viral proteins found in JEV-infected cells...
2015: PloS One
https://www.readbyqxmd.com/read/25694595/a-basic-cluster-in-the-n-terminus-of-yellow-fever-virus-ns2a-contributes-to-infectious-particle-production
#8
Stephanie Voßmann, Janett Wieseler, Romy Kerber, Beate Mareike Kümmerer
UNLABELLED: The flavivirus NS2A protein is involved in the assembly of infectious particles. To further understand its role in this process, a charged-to-alanine scanning analysis was performed on NS2A encoded by an infectious cDNA clone of yellow fever virus (YFV). Fifteen mutants containing single, double, or triple charged-to-alanine changes were tested. Five of them did not produce infectious particles, whereas efficient RNA replication was detectable for two of the five NS2A mutants (R22A-K23A-R24A and R99A-E100A-R101A mutants)...
May 2015: Journal of Virology
https://www.readbyqxmd.com/read/25653435/scanning-mutagenesis-studies-reveal-a-potential-intramolecular-interaction-within-the-c-terminal-half-of-dengue-virus-ns2a-involved-in-viral-rna-replication-and-virus-assembly-and-secretion
#9
Ren-Huang Wu, Ming-Han Tsai, Day-Yu Chao, Andrew Yueh
UNLABELLED: The NS2A protein of dengue virus (DENV) has eight predicted transmembrane segments (pTMSs; pTMS1 to pTMS8). NS2A has been shown to participate in RNA replication, virion assembly, and the host antiviral response. However, the role of the amino acid residues within the pTMS regions of NS2A during the virus life cycle is poorly understood. In the study described here, we explored the function of DENV NS2A by introducing a series of double or triple alanine substitutions into the C-terminal half (pTMS4 to pTMS8) of NS2A in the context of a DENV infectious clone or subgenomic replicon...
April 2015: Journal of Virology
https://www.readbyqxmd.com/read/25568208/characterization-of-dengue-virus-ns4a-and-ns4b-protein-interaction
#10
Jing Zou, Xuping Xie, Qing-Yin Wang, Hongping Dong, Michelle Yueqi Lee, Congbao Kang, Zhiming Yuan, Pei-Yong Shi
UNLABELLED: Flavivirus replication is mediated by a membrane-associated replication complex where viral membrane proteins NS2A, NS2B, NS4A, and NS4B serve as the scaffold for the replication complex formation. Here, we used dengue virus serotype 2 (DENV-2) as a model to characterize viral NS4A-NS4B interaction. NS4A interacts with NS4B in virus-infected cells and in cells transiently expressing NS4A and NS4B in the absence of other viral proteins. Recombinant NS4A and NS4B proteins directly bind to each other with an estimated Kd (dissociation constant) of 50 nM...
April 2015: Journal of Virology
https://www.readbyqxmd.com/read/25392211/two-distinct-sets-of-ns2a-molecules-are-responsible-for-dengue-virus-rna-synthesis-and-virion-assembly
#11
Xuping Xie, Jing Zou, Chunya Puttikhunt, Zhiming Yuan, Pei-Yong Shi
UNLABELLED: Flavivirus nonstructural protein 2A (NS2A) plays important roles in both viral RNA synthesis and virion assembly. The molecular details of how the NS2A protein modulates the two distinct events have not been defined. To address this question, we have performed a systematic mutagenesis of NS2A using dengue virus (DENV) serotype 2 (DENV-2) as a model. We identified two sets of NS2A mutations with distinct defects during a viral infection cycle. One set of NS2A mutations (D125A and G200A) selectively abolished viral RNA synthesis...
January 15, 2015: Journal of Virology
https://www.readbyqxmd.com/read/25375107/programmed-ribosomal-frameshift-alters-expression-of-west-nile-virus-genes-and-facilitates-virus-replication-in-birds-and-mosquitoes
#12
Ezequiel Balmori Melian, Sonja Hall-Mendelin, Fangyao Du, Nick Owens, Angela M Bosco-Lauth, Tomoko Nagasaki, Stephen Rudd, Aaron C Brault, Richard A Bowen, Roy A Hall, Andrew F van den Hurk, Alexander A Khromykh
West Nile virus (WNV) is a human pathogen of significant medical importance with close to 40,000 cases of encephalitis and more than 1,600 deaths reported in the US alone since its first emergence in New York in 1999. Previous studies identified a motif in the beginning of non-structural gene NS2A of encephalitic flaviviruses including WNV which induces programmed -1 ribosomal frameshift (PRF) resulting in production of an additional NS protein NS1'. We have previously demonstrated that mutant WNV with abolished PRF was attenuated in mice...
November 2014: PLoS Pathogens
https://www.readbyqxmd.com/read/24696348/construction-of-plasmid-bacterial-expression-purification-and-assay-of-dengue-virus-type-2-ns5-methyltransferase
#13
Siwaporn Boonyasuppayakorn, Radhakrishnan Padmanabhan
Dengue virus (DENV), a member of mosquito-borne flavivirus, causes self-limiting dengue fever as well as life-threatening dengue hemorrhagic fever and dengue shock syndrome. Its positive sense RNA genome has a cap at the 5'-end and no poly(A) tail at the 3'-end. The viral RNA encodes a single polyprotein, C-prM-E-NS1-NS2A-NS2B-NS3-NS4A-NS4B-NS5. The polyprotein is processed into 3 structural proteins (C, prM, and E) and 7 nonstructural (NS) proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5). NS3 and NS5 are multifunctional enzymes performing various tasks in viral life cycle...
2014: Methods in Molecular Biology
https://www.readbyqxmd.com/read/24696335/construction-of-self-replicating-subgenomic-dengue-virus-4-denv4-replicon
#14
Sofia L Alcaraz-Estrada, Rosa Del Angel, Radhakrishnan Padmanabhan
Dengue virus serotypes 1-4 are members of mosquito-borne flavivirus genus of Flaviviridae family that encode one long open reading frame (ORF) that is translated to a polyprotein. Both host and virally encoded proteases function in the processing of the polyprotein by co-translational and posttranslational mechanisms to yield 10 mature proteins prior to viral RNA replication. To study cis- and trans-acting factors involved in viral RNA replication, many groups [1-8] have constructed cDNAs encoding West Nile virus (WNV), DENV, or yellow fever virus reporter replicon RNAs...
2014: Methods in Molecular Biology
https://www.readbyqxmd.com/read/24316047/innate-immunity-to-dengue-virus-infection-and-subversion-of-antiviral-responses
#15
REVIEW
Angela M Green, P Robert Beatty, Alexandros Hadjilaou, Eva Harris
Dengue is a major public health issue in tropical and subtropical regions worldwide. The four serotypes of dengue virus (DENV1-DENV4) are spread primarily by Aedes aegypti and Aedes albopictus mosquitoes, whose geographic range continues to expand. Humans are the only host for epidemic strains of DENV, and the virus has developed sophisticated mechanisms to evade human innate immune responses. The host cell's first line of defense begins with an intracellular signaling cascade resulting in production of interferon α/β (IFN-α/β), which promotes intracellular antiviral responses and helps initiates the adaptive response during the course of DENV infection...
March 20, 2014: Journal of Molecular Biology
https://www.readbyqxmd.com/read/24074601/protein-protein-interactions-among-west-nile-non-structural-proteins-and-transmembrane-complex-formation-in-mammalian-cells
#16
Li Yu, Kazuyo Takeda, Lewis Markoff
To study the membrane orientation of flavivirus non-structural proteins (NSPs) in the replication complex, the seven major West Nile (WN) NSPs were separately expressed in monkey cells, and their subcellular localization was investigated by imaging-based techniques. First, we observed by confocal microscopy that four small transmembrane proteins (TP) (NS2A, NS2B, NS4A, and NS4B) were located to the endoplasmic reticulum (ER), whereas the largest NSPs, NS1, NS3, and NS5 were not. We then analyzed the colocalization and the association of WN NSPs using the methods of confocal microscopy, fluorescence resonance energy transfer (FRET), and biologic fluorescence complementation (BiFC)...
November 2013: Virology
https://www.readbyqxmd.com/read/23949785/molecular-characterization-of-a-duck-tembusu-virus-from-china
#17
XiaoFei Bai, Rang Lv, ChunGuo Liu, Na Qiu, Yilong He, XiuChen Yin, XiaoJun Li, Ming Liu, Yun Zhang
A new emerging flavivirus caused severe egg-drop in poultry and spread quickly across most duck-producing regions of China in 2010. Complete genome sequencing indicated that the virus genome is 10,989 nucleotides in length and possesses typical flavivirus genome organization, 5' untranslated region (UTR)-Cv-Ci-prM-M-E-NS1-NS2A-NS2B-NS3-NS4A-2K-NS4B-NS5-3'-UTR. The long open reading frame (ORF) encodes 3,425 amino acids (95-10,372 nt). The 94-nucleotide 5'-UTR is of intermediate size and the 617-nucleotide 3'-UTR is quite long relative to those of other flaviviruses...
December 2013: Virus Genes
https://www.readbyqxmd.com/read/23892127/simultaneous-detection-of-west-nile-and-japanese-encephalitis-virus-rna-by-duplex-taqman-rt-pcr
#18
Silvia C Barros, Fernanda Ramos, Líbia Zé-Zé, Maria J Alves, Teresa Fagulha, Margarida Duarte, Margarida Henriques, Tiago Luís, Miguel Fevereiro
West Nile virus (WNV) and Japanese encephalitis virus (JEV) are important mosquito-borne viruses of the Flaviviridae family, associated with encephalitis, mainly in humans and horses. WNV is also pathogen for many bird species. The incidence of human and animal WNV infections in Europe has risen, mostly in recent years, and JEV was detected in 2011 in mosquitoes collected in Italy and may emerge in Europe in the same way as other flaviviruses had emerged recently (USUTU and Bagaza virus) and should be regarded as a potential threat to public health...
November 2013: Journal of Virological Methods
https://www.readbyqxmd.com/read/23408612/membrane-topology-and-function-of-dengue-virus-ns2a-protein
#19
Xuping Xie, Shovanlal Gayen, CongBao Kang, Zhiming Yuan, Pei-Yong Shi
Flavivirus nonstructural protein 2A (NS2A) is a component of the viral replication complex that functions in virion assembly and antagonizes the host immune response. Although flavivirus NS2A is known to associate with the endoplasmic reticulum (ER) membrane, the detailed topology of this protein has not been determined. Here we report the first topology model of flavivirus NS2A on the ER membrane. Using dengue virus (DENV) NS2A as a model, we show that (i) the N-terminal 68 amino acids are located in the ER lumen, with one segment (amino acids 30 to 52) that interacts with ER membrane without traversing the lipid bilayer; (ii) amino acids 69 to 209 form five transmembrane segments, each of which integrally spans the ER membrane; and (iii) the C-terminal tail (amino acids 210 to 218) is located in the cytosol...
April 2013: Journal of Virology
https://www.readbyqxmd.com/read/23114626/west-nile-virus-ns2a-protein-facilitates-virus-induced-apoptosis-independently-of-interferon-response
#20
Ezequiel Balmori Melian, Judith H Edmonds, Tomoko Kim Nagasaki, Edward Hinzman, Nadia Floden, Alexander A Khromykh
The flavivirus NS2A protein is a small, multifunctional protein, involved in replication, virion formation and regulation of the innate immune response. Using the Kunjin strain of West Nile virus (WNV(KUN)) we previously demonstrated that a single amino acid change from alanine to proline at position 30 of the NS2A protein (A30P) reduced viral cytopathicity in cells and virulence in mice. To further investigate functions of the NS2A protein we have substituted alanine at position 30 with different amino acids (A30 mutants) in a WNV(KUN) infectious clone...
February 2013: Journal of General Virology
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