Abstract:
:Recombinant viruses possessing reporter proteins have been generated for virus research. In the case of the family Flaviviridae, we recently generated recombinant viruses, including the hepatitis C virus of the genus Hepacivirus, Japanese encephalitis virus (JEV) of the genus Flavivirus, and bovine viral diarrhea virus of the genus Pestivirus; all three viruses possess an 11-amino-acid subunit derived from NanoLuc luciferase (HiBiT). Here, we further developed the recombinant viruses and investigated their utility in vivo Recombinant viruses harboring HiBiT in the E, NS1, or NS3 protein constructed based on the predicted secondary structure, solvent-accessible surface area, and root mean square fluctuation of the proteins exhibited comparable replication to that of the wild-type virus in vitro The recombinant JEV carrying HiBiT in the NS1 protein exhibited propagation in mice comparable to that of the parental virus, and propagation of the recombinant was monitored by the luciferase activity. In addition, the recombinants of classical swine fever virus (CSFV) possessing HiBiT in the Erns or E2 protein also showed propagation comparable to that of the wild-type virus. The recombinant CSFV carrying HiBiT in Erns exhibited similar replication to the parental CSFV in pigs, and detection of viral propagation of this recombinant by luciferase activity was higher than that by quantitative PCR (qPCR). Taken together, these results demonstrated that the reporter Flaviviridae viruses generated herein are powerful tools for elucidating the viral life cycle and pathogeneses and provide a robust platform for the development of novel antivirals.IMPORTANCEIn vivo applications of reporter viruses are necessary to understand viral pathogenesis and provide a robust platform for antiviral development. In developing such applications, determination of an ideal locus to accommodate foreign genes is important, because insertion of foreign genes into irrelevant loci can disrupt the protein functions required for viral replication. Here, we investigated the criteria to determine ideal insertion sites of foreign genes from the protein structure of viral proteins. The recombinant viruses generated by our criteria exhibited propagation comparable to that of parental viruses in vivo Our proteomic approach based on the flexibility profile of viral proteins may provide a useful tool for constructing reporter viruses, including Flaviviridae viruses.
journal_name
J Viroljournal_title
Journal of virologyauthors
Tamura T,Igarashi M,Enkhbold B,Suzuki T,Okamatsu M,Ono C,Mori H,Izumi T,Sato A,Fauzyah Y,Okamoto T,Sakoda Y,Fukuhara T,Matsuura Ydoi
10.1128/JVI.01191-19subject
Has Abstractpub_date
2019-10-29 00:00:00issue
22eissn
0022-538Xissn
1098-5514pii
JVI.01191-19journal_volume
93pub_type
杂志文章abstract::Linear hepadnavirus DNA in primary hepatocyte cultures efficiently participates in intra- and intermolecular nonhomologous recombination at its ends. The products of this recombination are (i) monomeric covalently closed circular DNAs (cccDNAs) with deletions and insertions around the site of joining and (ii) oligomer...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.7.4029-4036.1995
更新日期:1995-07-01 00:00:00
abstract::The mechanism by which murine polyomavirus penetrates cells and arrives at the nucleus, the site of viral replication, is not well understood. Simian virus 40 and JC virus, two closely related members of the polyomavirus subfamily, use caveola- and clathrin-mediated uptake pathways for entry, respectively. The data pr...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.18.8582-8588.2000
更新日期:2000-09-01 00:00:00
abstract::Simian virus 40 (SV40) infection of human diploid cells failed to cause an enhanced production of thymidine kinase during the first 10 days after infection. Thymidine kinase activities from extracts of SV40-transformed cultures (human or simian) were considerably higher than the activity levels in extracts from the no...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.1.5.912-919.1967
更新日期:1967-10-01 00:00:00
abstract::Previous studies have indicated that B cells make a significant contribution to the resolution of influenza virus infection. To determine how B cells participate in the control of the infection, we transferred intact, major histocompatibility complex class II (MHC-II)-negative or B-cell receptor (BCR)-transgenic splee...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.10.5943-5951.2005
更新日期:2005-05-01 00:00:00
abstract::Here, we report that specific manipulations of the cellular response to virus infection can cause prevention of apoptosis and consequent establishment of persistent infection. Infection of several human cell lines with Sendai virus (SeV) or human parainfluenza virus 3, two prototypic paramyxoviruses, caused slow apopt...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02536-07
更新日期:2008-04-01 00:00:00
abstract::Translation of aphthovirus RNA is initiated at an internal ribosome entry site (IRES) element, preceding the first functional AUG initiation codon. The effect of mutations at the base of domain 3 of the aphthovirus IRES on translation activity has been analyzed by site-directed mutagenesis and expression of bicistroni...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.70.2.992-998.1996
更新日期:1996-02-01 00:00:00
abstract::Waterfowl are the natural reservoir of all influenza A viruses, which are usually nonpathogenic in wild aquatic birds. However, in late 2002, outbreaks of highly pathogenic H5N1 influenza virus caused deaths among wild migratory birds and resident waterfowl, including ducks, in two Hong Kong parks. In February 2003, a...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.78.9.4892-4901.2004
更新日期:2004-05-01 00:00:00
abstract::Positional homologs to the UL51 open reading frame of herpes simplex virus type 1 have been identified throughout the herpesvirus family. However, no respective protein has so far been described for any of the herpesviruses. With rabbit antisera directed against oligopeptides predicted to comprise antigenic regions of...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.7.5635-5638.1997
更新日期:1997-07-01 00:00:00
abstract::When mouse L cells are infected for 22 hr with vesicular stomatitis virus (VSV), a ribonucleic acid-containing enveloped virus, greater than 70% of the major histocompatibility antigen (H-2), is no longer detectable by the method of inhibition of immune cytolysis. Infected cells prelabeled with (14)C-glucosamine also ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.10.4.578-585.1972
更新日期:1972-10-01 00:00:00
abstract::Parapoxviruses can be morphologically distinguished from other poxviruses in conventional negative staining electron microscopy (EM) by their ovoid appearance and the spiral tubule surrounding the virion's surface. However, this technique may introduce artifacts. We have examined Orf virus (ORFV; the prototype species...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.78.15.8085-8093.2004
更新日期:2004-08-01 00:00:00
abstract:UNLABELLED:Transmission of chronic wasting disease (CWD) between cervids is influenced by the primary structure of the host cellular prion protein (PrP(C)). In white-tailed deer, PRNP alleles encode the polymorphisms Q95 G96 (wild type [wt]), Q95 S96 (referred to as the S96 allele), and H95 G96 (referred to as the H95 ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02010-15
更新日期:2015-12-01 00:00:00
abstract::Activation of the interferon-inducible, double-stranded RNA-dependent protein kinase was monitored in monolayer cultures of control and interferon-treated HeLa cells infected with encephalomyocarditis virus. The extent of phosphorylation in the intact cell of the alpha-subunit of eucaryotic protein synthesis initiatio...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.54.3.894-898.1985
更新日期:1985-06-01 00:00:00
abstract::The vaccinia virus H2R gene (VACWR 100) is conserved in all sequenced members of the poxvirus family and encodes a protein with a predicted transmembrane domain and four invariant cysteines. A recombinant vaccinia virus, in which expression of the H2 protein is stringently regulated, was unable to replicate without in...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.8.4744-4754.2005
更新日期:2005-04-01 00:00:00
abstract::Reovirus attachment protein σ1 engages glycan receptors and junctional adhesion molecule-A (JAM-A) and is thought to undergo a conformational change during the proteolytic disassembly of virions to infectious subvirion particles (ISVPs) that accompanies cell entry. The σ1 protein is also the primary target of neutrali...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01621-16
更新日期:2017-01-31 00:00:00
abstract::Artificial target sequences for tissue-specific miRNAs have recently been introduced as a new means for altering the tissue tropism of viral replication. This approach can be used to improve the safety of oncolytic viruses for cancer virotherapy by restricting their replication in unwanted tissues, such as the liver. ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01940-12
更新日期:2013-01-01 00:00:00
abstract::Processes by which human herpesviruses penetrate and are released from polarized epithelial cells, which have distinct apical and basolateral membrane domains differing in protein and lipid content, are poorly understood. We recently reported that human cytomegalovirus (CMV) mutants with deletions of the gene US9 form...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.72.7.5717-5727.1998
更新日期:1998-07-01 00:00:00
abstract::The promoter of the murine thymidine kinase gene contains a binding site for transcription factor E2F. Using cell lines (3T3-LT) conditionally expressing polyomavirus large T antigen from a hormone-responsive promoter and reporter gene constructs carrying the thymidine kinase promoter with intact or mutated E2F sites,...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.67.4.1765-1771.1993
更新日期:1993-04-01 00:00:00
abstract::We reported previously that the movement protein (MP) of tomato mosaic tobamovirus is phosphorylated, and we proposed that MP phosphorylation is important for viral pathogenesis. Experimental data indicated that phosphorylation enhances the stability of MP in vivo and enables the protein to assume the correct intracel...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.77.2.1452-1461.2003
更新日期:2003-01-01 00:00:00
abstract::Two major outer envelope glycoproteins of Epstein-Barr virus, gp350 and gp220, are known to be encoded by 3.2- and 2.5-kilobase RNAs which map to the same DNA fragment (M. Hummel, D. Thorley-Lawson, and E. Kieff, J. Virol. 49:413-417). These RNAs have the same 5' and 3' ends. The larger RNA is encoded by a 2,777-base ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.54.3.665-674.1985
更新日期:1985-06-01 00:00:00
abstract::Transmission of human immunodeficiency virus type 1 (HIV-1) occurs primarily via the mucosal route, suggesting that HIV-1 vaccines may need to elicit mucosal immune responses. Here, we investigated the immunogenicity and relative efficacy of systemic immunization with two human ALVAC-HIV-1 recombinant vaccines express...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.80.8.3732-3742.2006
更新日期:2006-04-01 00:00:00
abstract::A general theme that has emerged from studies of DNA tumor viruses is that otherwise unrelated oncoproteins encoded by these viruses often target the same important cellular factors. Major oncogenic determinants for human adenovirus type 9 (Ad9) and high-risk human papillomaviruses (HPV) are the E4-ORF1 and E6 oncopro...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.20.9680-9693.2000
更新日期:2000-10-01 00:00:00
abstract::The N termini of the capsid proteins VP1 and VP2 of adeno-associated virus (AAV) play important roles in subcellular steps of infection and contain motifs that are highly homologous to a phospholipase A(2) (PLA(2)) domain and nuclear localization signals (NLSs). To more clearly understand how virion components influen...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00687-10
更新日期:2010-09-01 00:00:00
abstract::A cloned segment of the polyoma virus genome encoding the small T antigen has been fused, in the correct phase for translation, to the 5' end of the beta-galactosidase gene. The hybrid gene, cloned in Escherichia coli, produces a protein resembling the small T antigen. ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.36.1.125-132.1980
更新日期:1980-10-01 00:00:00
abstract::A specific endoribonucleolytic activity was detected when detergent-lysed vesicular stomatitis of Sendai virus was incubated with the precursor to Escherichia coli tRNA Tyr. The cleavage products produced and the characteristics of the reaction were similar to those previously reported for human KB cell RNase NU. Like...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.25.1.274-284.1978
更新日期:1978-01-01 00:00:00
abstract::HIV protease is known to cause cell death, which is dependent upon cleavage of procaspase 8. HIV protease cleavage of procaspase 8 generates Casp8p41, which directly binds Bak with nanomolar affinity, causing Bak activation and consequent cell death. Casp8p41 can also bind Bcl2 with nanomolar affinity, in which case c...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00037-18
更新日期:2018-06-13 00:00:00
abstract::Ebola viruses (EboV) are enveloped RNA viruses infecting cells by a pH-dependent process mediated by viral glycoproteins (GP) involving endocytosis of virions and their routing into acidic endosomes. As with well-characterized pH-dependent viral entry proteins, in particular influenza virus hemagglutinin, it is though...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.80.6.2815-2822.2006
更新日期:2006-03-01 00:00:00
abstract::We have evaluated the potential of conferring protective immunity to herpes simplex virus type 2 (HSV-2) by selectively inducing an HSV-specific CD8(+) cytotoxic T-lymphocyte (CTL) response directed against a single major histocompatibility complex class I-restricted CTL recognition epitope. We generated a recombinant...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.72.12.9567-9574.1998
更新日期:1998-12-01 00:00:00
abstract::The glucocorticoid-regulatory sequences from the murine mammary tumor virus long terminal repeat (MMTV LTR) were introduced into the LTR of Moloney murine leukemia virus (M-MuLV) by recombinant DNA techniques. The site of insertion was in the M-MuLV LTR U3 region at -150 base pairs with respect to the RNA cap site. In...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.54.1.133-144.1985
更新日期:1985-04-01 00:00:00
abstract::The goose parvovirus (GPV) Rep 1 and Rep 2 proteins are encoded by P9-generated mRNAs that are either unspliced or spliced within the rep gene region, respectively. These mRNAs are present in an approximately equal ratio. The translation of Rep 1 was initiated from the first AUG in unspliced P9-generated mRNA; however...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01255-09
更新日期:2009-10-01 00:00:00
abstract::The 2009 H1N1 influenza pandemic provided an opportunity to study human virus-specific T cell responses after infection with a novel influenza virus against which limited humoral immunity existed in the population. Here we describe the magnitude, kinetics, and nature of the virus-specific T cell response using intrace...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.05204-11
更新日期:2011-11-01 00:00:00