Abstract:
:Engineered recombinant viruses expressing reporter genes have been developed for real-time monitoring of replication and for mass screening of antiviral inhibitors. Recently, we reported using a reverse genetics system to develop the first recombinant reporter rotaviruses (RVs) that expressed NanoLuc (NLuc) luciferase. Here, we describe a strategy for developing stable reporter RVs expressing luciferase and green or red fluorescent proteins. The reporter genes were inserted into the open reading frame of NSP1 and expressed as a fusion with an NSP1 peptide consisting of amino acids 1 to 27. The stability of foreign genes within the reporter RV strains harboring a shorter chimeric NSP1-reporter gene was greater than that of those in the original reporter RV strain, independent of the transgene inserted. The improved reporter RV was used to screen for neutralizing monoclonal antibodies (MAbs). Sequence analysis of escape mutants from one MAb clone (clone 29) identified an amino acid substitution (arginine to glycine) at position 441 in the VP4 protein, which resides within neutralizing epitope 5-1 in the VP5* fragment. Furthermore, to express a native reporter protein lacking NSP1 amino acids 1 to 27, the 5'- and 3'-terminal region sequences were modified to restore the predicted secondary RNA structure of the NSP1-reporter chimeric gene. These data demonstrate the utility of reporter RVs for live monitoring of RV infections and also suggest further applications (e.g., RV vaccine vectors, which can induce mucosal immunity against intestinal pathogens).IMPORTANCE Development of reporter RVs has been hampered by the lack of comprehensive reverse genetics systems. Recently, we developed a plasmid-based reverse genetics system that enables generation of reporter RVs expressing NLuc luciferase. The prototype reporter RV had some disadvantages (i.e., the transgene was unstable and was expressed as a fusion protein with a partial NSP1 peptide); however, the improved reporter RV overcomes these problems through modification of the untranslated region of the reporter-NSP1 chimeric gene. This strategy for generating stable reporter RVs could be expanded to diverse transgenes and be used to develop RV transduction vectors. Also, the data improve our understanding of the importance of 5'- and 3'-terminal sequences in terms of genome replication, assembly, and packaging.
journal_name
J Viroljournal_title
Journal of virologyauthors
Kanai Y,Kawagishi T,Nouda R,Onishi M,Pannacha P,Nurdin JA,Nomura K,Matsuura Y,Kobayashi Tdoi
10.1128/JVI.01774-18subject
Has Abstractpub_date
2019-02-05 00:00:00issue
4eissn
0022-538Xissn
1098-5514pii
JVI.01774-18journal_volume
93pub_type
杂志文章abstract::The configuration of latent varicella-zoster virus (VZV) DNA was analyzed by PCR. Template DNA for both internal and terminal VZV primers was present in a 1:1 ratio in ganglionic DNA, compared with a 15:1 ratio in DNA extracted from VZV virions, indicating that the VZV genomic termini are adjacent in latently infected...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.12.8151-8154.1995
更新日期:1995-12-01 00:00:00
abstract::Mature dendritic cells (DCs) are potent antigen-presenting cells essential for initiating successful antiviral immune responses and would therefore serve as an ideal target for viruses seeking to evade or delay the immune response by disrupting their function. We have previously reported that VZV productively infects ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.77.8.4950-4959.2003
更新日期:2003-04-01 00:00:00
abstract::The large T antigen encoded by simian virus 40 (SV40) plays essential roles in the infection of permissive cells, leading to production of progeny virions, and in the infection of nonpermissive cells, leading to malignant transformation. Primary mouse embryo fibroblasts (MEFs) are nonpermissive for SV40, and infection...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.65.12.6872-6880.1991
更新日期:1991-12-01 00:00:00
abstract::Human rhinoviruses (HRVs) are a highly prevalent and diverse group of respiratory viruses. Although HRV-A and HRV-B are traditionally detected by virus isolation, a series of unculturable HRV variants have recently been described and assigned as a new species (HRV-C) within the picornavirus Enterovirus genus. To inves...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00962-10
更新日期:2010-10-01 00:00:00
abstract::The retrovirus SL3 induces T-cell lymphomas in mice. The transcriptional enhancer in the long terminal repeat (LTR) of SL3 contains two 72-bp repeats. Each repeat contains a binding site for the transcription factor CBF (also called AML1). The CBF binding sites are called core elements. SAA is a mutant that is identic...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.73.9.7599-7606.1999
更新日期:1999-09-01 00:00:00
abstract::A hallmark of infection with herpes simplex virus type 1 (HSV-1) is the establishment of latency in ganglia of the infected individual. During the life of the latently infected individual, the virus can occasionally reactivate, travel back to the eye, and cause recurrent disease. Indeed, a major cause of corneal scarr...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02234-08
更新日期:2009-03-01 00:00:00
abstract::Previous experiments have demonstrated that adenovirus infection of human and mouse cells leads to an E1A-dependent activation of the DNA-binding capacity of a cellular transcription factor termed E2F. E2F binds to two sites in the adenovirus E2 early promoter which have been shown to be critical for E1A-dependent E2 ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.63.9.3643-3650.1989
更新日期:1989-09-01 00:00:00
abstract::Alternative splicing of adeno-associated virus type 2 (AAV2) P19-generated pre-mRNAs generates the small Rep proteins Rep52 and Rep40, which differ in their carboxyl termini. Both proteins are required for optimal packaging of AAV2 genomes. AAV5 Rep-encoding P19-generated transcripts are primarily polyadenylated withi...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01961-09
更新日期:2010-01-01 00:00:00
abstract::The involvement of the mouse mammary tumor virus (MTV) in spontaneous and hormone-induced mammary tumors in low-mammary-tumor mouse strains was studied by comparing the amounts of MTV RNA and MTV DNA sequences in mammary tumors and other tissues of mice with an without hormonal treatments. The following results were o...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.27.3.551-559.1978
更新日期:1978-09-01 00:00:00
abstract::The envelope glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV) is posttranslationally cleaved into two subunits. We show here that this endoproteolytic processing is not required for transport to the cell surface but is essential for LCMV GP to mediate infectivity of pseudotyped retroviral vectors. By sys...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.77.5.2866-2872.2003
更新日期:2003-03-01 00:00:00
abstract::A technique of continuous filtration of T4-infected Escherichia coli has been devised to study the phenomenon of lysis inhibition. Studies using this technique revealed that the length of the lysis delay caused by superinfection can attain only certain discrete values, which for low average multiplicity of superinfect...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.1.5.948-955.1967
更新日期:1967-10-01 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::The human cyclin T1 (hCycT1) protein from the positive transcription elongation factor b (P-TEFb) binds the transactivator Tat and the transactivation response (TAR) RNA stem loop from human immunodeficiency virus type 1 (HIV). This complex activates the elongation of viral transcription. To create effective inhibitor...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.76.21.10873-10881.2002
更新日期:2002-11-01 00:00:00
abstract::Breast cancer is the second leading cause of cancer-related deaths in women in the United States. Triple-negative breast cancer constitutes a subset of breast cancer that is associated with higher rates of relapse, decreased survival, and limited therapeutic options for patients afflicted with this type of breast canc...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01411-19
更新日期:2019-11-13 00:00:00
abstract::Cholesterol has been implicated in various viral life cycle steps for different enveloped viruses, including viral entry into host cells, cell-cell fusion, and viral budding from infected cells. Enveloped viruses acquire their membranes from their host cells. Though cholesterol has been associated with binding and ent...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02323-20
更新日期:2021-01-06 00:00:00
abstract::Hepatitis C virus (HCV) is an important human pathogen affecting an estimated 3% of the world's population. Recent advances have enabled in vitro propagation of the virus and allow assembly and egress to be investigated for the first time. As a component of the virion, the HCV core protein likely functions primarily i...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00793-07
更新日期:2007-10-01 00:00:00
abstract::Effectively recognizing invading viruses and subsequently inducing innate antiviral immunity are essential for host antiviral defense. Although these processes are closely regulated by the host to maintain immune balance, viruses have evolved the ability to downregulate or upregulate these processes for their survival...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01594-17
更新日期:2018-01-02 00:00:00
abstract:UNLABELLED:Susceptibility to alphavirus infection is age dependent, and host maturation is associated with decreased virus replication and less severe encephalitis. To identify factors associated with maturation-dependent restriction of virus replication, we studied AP-7 rat olfactory bulb neuronal cells, which can dif...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02394-14
更新日期:2015-01-01 00:00:00
abstract::Arginine methylation of human immunodeficiency virus type 1 (HIV-1) Tat protein downregulates its key function in viral-gene transactivation. The fate of methylated Tat is unknown, so it is unclear whether methylated Tat is degraded or persists in the cell for additional functions. Here we show that the arginine methy...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00499-09
更新日期:2009-11-01 00:00:00
abstract::Replication of the human herpesvirus Epstein-Barr virus drastically impairs cellular protein synthesis. This shutoff phenotype results from mRNA degradation upon expression of the early lytic-phase protein BGLF5. Interestingly, BGLF5 is the viral DNase, or alkaline exonuclease, homologues of which are present througho...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00309-12
更新日期:2012-09-01 00:00:00
abstract::A continuous cell line of highly contact-inhibited cells (NIH/3T3) has been developed from NIH Swiss mouse embryo cultures. Its growth properties are similar to those of 3T3 and BALB/3T3. Although 3T3 is relatively insensitive to focus formation by murine sarcoma viruses, cloned lines of both NIH/3T3 and BALB/3T3 have...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.4.5.549-553.1969
更新日期:1969-11-01 00:00:00
abstract::The phenotypic properties of ICP27 temperature-sensitive and deletion mutants and the results of transient expression assays have demonstrated that ICP27 has a modulatory effect on viral gene expression induced by ICPs 0 and 4. In order to identify the regions of the ICP27 molecule that are responsible for its enhanci...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.64.7.3471-3485.1990
更新日期:1990-07-01 00:00:00
abstract::The RNA-dependent RNA polymerase of influenza A virus is responsible for both transcription and replication of negative-sense viral RNA. It is thought that a "switching" mechanism regulates the transition between these activities. We demonstrate that, in the presence of preexisting viral RNA polymerase and nucleoprote...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.78.17.9568-9572.2004
更新日期:2004-09-01 00:00:00
abstract::In a previous study (S. Zheng, G. N. Woode, D. R. Melendy, and R. F. Ramig, J. Clin. Microbiol. 27:1939-1945, 1989), it was predicted that the VP7 serotype 6 bovine rotavirus strains NCDV and B641 do not share antigenically similar VP4s. In this study, gene 4 and the VP7 gene of B641 were sequenced, and the amino acid...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.65.10.5535-5538.1991
更新日期:1991-10-01 00:00:00
abstract::Candidate foot-and-mouth disease (FMD) DNA vaccines designed to produce viral capsids lacking infectious viral nucleic acid were evaluated. Plasmid DNAs containing a portion of the FMDV genome coding for the capsid precursor protein (P1-2A) and wild-type or mutant viral proteinase 3C (plasmids P12X3C or P12X3C-mut, re...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.72.5.4454-4457.1998
更新日期:1998-05-01 00:00:00
abstract::Hydroxyurea, an inhibitor of ribonucleotide reductase, blocks replication of vaccinia virus. However, when medium containing hydroxyurea and dialyzed serum was supplemented with deoxyadenosine, the block to viral reproduction was circumvented, provided that an inhibitor of adenosine deaminase was also present. Deoxygu...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.65.5.2290-2298.1991
更新日期:1991-05-01 00:00:00
abstract::We analyzed a region of the capsid of canine parvovirus (CPV) which determines the ability of the virus to infect canine cells. This region is distinct from those previously shown to determine the canine host range differences between CPV and feline panleukopenia virus. It lies on a ridge of the threefold spike of the...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.12.9214-9222.1997
更新日期:1997-12-01 00:00:00
abstract::The functions of bovine respiratory syncytial virus (BRSV) nonstructural proteins NS1 and NS2 were studied by generation and analysis of recombinant BRSV carrying single and double gene deletions. Whereas in MDBK cells the lack of either or both NS genes resulted in a 5,000- to 10,000-fold reduction of virus titers, i...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.18.8234-8242.2000
更新日期:2000-09-01 00:00:00
abstract::Defined segments of the transmembrane envelope glycoprotein (gp45) of equine infectious anemia virus were expressed as TrpLE fusion proteins and examined for their reactivity in Western immunoblots against a diverse panel of equine immune sera. The most immunogenic region of gp45 was localized to its amino terminus, p...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.65.2.1013-1018.1991
更新日期:1991-02-01 00:00:00
abstract::Vaccine-elicited humoral immune responses comprise an array of antibody forms and specificities, with only a fraction contributing to protective host immunity. Elucidation of antibody effector functions responsible for protective immunity against human immunodeficiency virus type 1 (HIV-1) acquisition is a major goal ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01552-17
更新日期:2018-03-14 00:00:00