NDLI: Rescue of an influenza A virus wild-type PB2 gene and a mutant derivative bearing a site-specific temperature-sensitive and attenuating mutation.

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Picornavirus nonstructural proteins: emerging roles in virus replication and inhibition of host cell functions.

Induction of gene expression by exon 2 of the major E1A proteins of adenovirus type 5.

Efficient self-assembly of human papillomavirus type 16 L1 and L1-L2 into virus-like particles.

Increased spacing between Sp1 and TATAA renders human immunodeficiency virus type 1 replication defective: implication for Tat function.

Renaturation of complementary DNA strands by herpes simplex virus type 1 ICP8.

Multiple double-stranded RNA segments are associated with virus particles infecting Trichomonas vaginalis.

Effects of the tat and nef gene products of human immunodeficiency virus type 1 (HIV-1) on transcription controlled by the HIV-1 long terminal repeat and on cell growth in macrophages.

The RNAs of hepatitis delta virus are copied by RNA polymerase II in nuclear homogenates.

Patterns of frog virus 3 DNA methylation and DNA methyltransferase activity in nuclei of infected cells.

Eleven loci encoding trans-acting factors are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA replication.

Superinfection-induced apoptosis and its correlation with the reduction of viral progeny in cells persistently infected with Hz-1 baculovirus.

Internal cleavage and trans-proteolytic activities of the VPg-proteinase (NIa) of tobacco etch potyvirus in vivo.

Human T-cell leukemia virus type 1 Tax activates transcription of the human fra-1 gene through multiple cis elements responsive to transmembrane signals.

T-cell activation influences initial DNA synthesis of simian immunodeficiency virus in resting T lymphocytes from macaques.

Aleutian mink disease parvovirus infection of mink macrophages and human macrophage cell line U937: demonstration of antibody-dependent enhancement of infection.

Induction of reactivation of herpes simplex virus in murine sensory ganglia in vivo by cadmium.

Dysregulated surface gene expression from disrupted hepatitis B virus genomes.

Characterization of the Ld-restricted cytotoxic T-lymphocyte epitope in the mouse hepatitis virus nucleocapsid protein.

Characterization of mouse hepatitis virus-specific cytotoxic T cells derived from the central nervous system of mice infected with the JHM strain.

Characterization of stable Chinese hamster ovary cells expressing wild-type, secreted, and glycosylphosphatidylinositol-anchored human immunodeficiency virus type 1 envelope glycoprotein.

Conditional infectivity of a human immunodeficiency virus matrix domain deletion mutant.

Influence of substrate structure on disintegration activity of Moloney murine leukemia virus integrase.

Pathogenicity of molecularly cloned bovine leukemia virus.

5' sequences of rubella virus RNA stimulate translation of chimeric RNAs and specifically interact with two host-encoded proteins.

Amplification and sequence analysis of DNA flanking integrated proviruses by a simple two-step polymerase chain reaction method.

Mutated primer binding sites interacting with different tRNAs allow efficient murine leukemia virus replication.

Two E2 binding sites (E2BS) alone or one E2BS plus an A/T-rich region are minimal requirements for the replication of the human papillomavirus type 11 origin.

Escape from in vivo restriction of Moloney mink cell focus-inducing viruses driven by the Mo+PyF101 long terminal repeat (LTR) by LTR alterations.

Isolation of a herpes simplex virus type 1 mutant with a deletion in the virion host shutoff gene and identification of multiple forms of the vhs (UL41) polypeptide.

Heterologous C-terminal sequences disrupt transcriptional activation and oncogenesis by p59v-rel.

Viral genomes maintained extrachromosomally in hamster polyomavirus-induced lymphomas display a cell-specific replication in vitro.

Bromovirus RNA replication and transcription require compatibility between the polymerase- and helicase-like viral RNA synthesis proteins.

Specific binding of human immunodeficiency virus type 1 gag polyprotein and nucleocapsid protein to viral RNAs detected by RNA mobility shift assays.

Mutually exclusive binding of two cellular factors within a critical promoter region of the gene for the IE110k protein of herpes simplex virus.

Three different cellular proteins bind to complementary sites on the 5'-end-positive and 3'-end-negative strands of mouse hepatitis virus RNA.

Rescue of an influenza A virus wild-type PB2 gene and a mutant derivative bearing a site-specific temperature-sensitive and attenuating mutation.

Incorporation of Vpr into human immunodeficiency virus type 1 virions: requirement for the p6 region of gag and mutational analysis.

Vpu-induced degradation of CD4: requirement for specific amino acid residues in the cytoplasmic domain of CD4.

Vesicular stomatitis virus M protein may be inside the ribonucleocapsid coil.

Binding sites for the herpes simplex virus immediate-early protein ICP4 impose an increased dependence on viral DNA replication on simple model promoters located in the viral genome.

Purification of the late transcription system of vaccinia virus: identification of a novel transcription factor.

Human immunodeficiency virus type 1 pol gene mutations in an AIDS patient treated with multiple antiretroviral drugs.

Herpes simplex virus type 1 latency-associated transcript (LAT) promoter deletion mutants can express a 2-kilobase transcript mapping to the LAT region.

Poliovirus capsid proteins derived from P1 precursors with glutamine-valine cleavage sites have defects in assembly and RNA encapsidation.

Epstein-Barr virus recombinants from overlapping cosmid fragments.

Transcriptional activation of the human T-lymphotropic virus type I long terminal repeat by functional interaction of Tax1 and Ets1.

Identification of the activation domain of equine infectious anemia virus rev.

Individual repeats of Drosophila Myb can function in transformation by v-Myb.

Molecular determinants of macrophage tropism and viral persistence: importance of single amino acid changes in the polymerase and glycoprotein of lymphocytic choriomeningitis virus.

Replication of lymphocytic choriomeningitis virus is restricted in terminally differentiated neurons.

Herpesvirus proteinase: site-directed mutagenesis used to study maturational, release, and inactivation cleavage sites of precursor and to identify a possible catalytic site serine and histidine.

A novel class of transcripts expressed with late kinetics in the absence of ICP4 spans the junction between the long and short segments of the herpes simplex virus type 1 genome.

Conformational changes induced in the envelope glycoproteins of the human and simian immunodeficiency viruses by soluble receptor binding.

Disulfide bonds in folding and transport of mouse hepatitis coronavirus glycoproteins.

Sequences determining the pH dependence of viral entry are distinct from the host range-determining region of the murine ecotropic and amphotropic retrovirus envelope proteins.

Epstein-Barr virus recombinants with specifically mutated BCRF1 genes.

Hepadnavirus infection requires interaction between the viral pre-S domain and a specific hepatocellular receptor.

High viral load in lymph nodes and latent human immunodeficiency virus (HIV) in peripheral blood cells of HIV-1-infected chimpanzees.

HLA A2.1-restricted cytotoxic T cells recognizing a range of Epstein-Barr virus isolates through a defined epitope in latent membrane protein LMP2.

The hemagglutinins of duck and human H1 influenza viruses differ in sequence conservation and in glycosylation.

Interleukin-2 downregulates hepatitis B virus gene expression in transgenic mice by a posttranscriptional mechanism.

In vivo footprinting analysis of constitutive and inducible protein-DNA interactions at the long terminal repeat of human immunodeficiency virus type 1.

Minimum internal ribosome entry site required for poliovirus infectivity.

Comparing regions of the Epstein-Barr virus ZEBRA protein which function as transcriptional activating sequences in Saccharomyces cerevisiae and in B cells.

Immune escape by human immunodeficiency virus type 1 from neutralizing antibodies: evidence for multiple pathways.

The herpes simplex virus type 1 regulatory protein ICP0 enhances virus replication during acute infection and reactivation from latency.

Divergence and evolution of homologous regions of Bombyx mori nuclear polyhedrosis virus.

Hepatitis C virus (HCV)-specific cytotoxic T lymphocytes recognize epitopes in the core and envelope proteins of HCV.

Dynamic equilibrium between vesicular stomatitis virus glycoprotein monomers and trimers in the Golgi and at the cell surface.

Human cytomegalovirus IE86 protein interacts with promoter-bound TATA-binding protein via a specific region distinct from the autorepression domain.

Site-specific inhibition of RNA polymerase II preinitiation complex assembly by human cytomegalovirus IE86 protein.

Adenovirus E4orf4 protein binds to protein phosphatase 2A, and the complex down regulates E1A-enhanced junB transcription.

Vaccinia virus nucleoside triphosphate phosphohydrolase I controls early and late gene expression by regulating the rate of transcription.

Varicella-zoster virus DNA polymerase and major DNA-binding protein genes have overlapping divergent promoters.

Reverse transcriptase and protease activities of avian leukosis virus Gag-Pol fusion proteins expressed in insect cells.

Role of spike protein conformational changes in fusion of Semliki Forest virus.

The DNA-binding domain of simian virus 40 tumor antigen has multiple functions.

NF-kappa B p100 is one of the high-molecular-weight proteins complexed with the v-Rel oncoprotein in transformed chicken spleen cells.

F0-containing noninfectious Sendai virus can initiate replication in mouse lungs but requires a relatively long incubation period.

Effects of virally expressed interleukin-10 on vaccinia virus infection in mice.

In vivo depletion of CD8+ T cells prevents lesions of demyelination in Semliki Forest virus infection.

Mutants of Epstein-Barr virus with a selective marker disrupting the TP gene transform B cells and replicate normally in culture.

Analysis of p107-associated proteins: p107 associates with a form of E2F that differs from pRB-associated E2F-1.

Functional interaction between Epstein-Barr virus DNA polymerase catalytic subunit and its accessory subunit in vitro.

Epstein-Barr virus induces fragmentation of chromosomal DNA during lytic infection.

Isoprenylation mediates direct protein-protein interactions between hepatitis large delta antigen and hepatitis B virus surface antigen.

Identification and characterization of an extended Tax protein in human T-cell lymphotropic virus type II subtype b isolates.

Interconnection between thyroid hormone signalling pathways and parvovirus cytotoxic functions.

Immunization with the immediate-early tegument protein (open reading frame 62) of varicella-zoster virus protects guinea pigs against virus challenge.

Identification of a lytic-phase origin of DNA replication in human herpesvirus 6B strain Z29.

Multiple, distinct trans-activation functions are encoded by the simian virus 40 large T and small t antigens, only some of which require the 82-residue amino-terminal common domain.

Mouse mammary tumor virus-induced tumorigenesis in sag transgenic mice: a laboratory model of natural selection.

Mechanism of interferon action: evidence for intermolecular autophosphorylation and autoactivation of the interferon-induced, RNA-dependent protein kinase PKR.

The human T-cell leukemia/lymphotropic virus type I p12I protein cooperates with the E5 oncoprotein of bovine papillomavirus in cell transformation and binds the 16-kilodalton subunit of the vacuolar H+ ATPase.

Hierarchy of polyadenylation site usage by bovine papillomavirus in transformed mouse cells.

Molecular mimicry between the human immunodeficiency virus type 1 gp120 V3 loop and human brain proteins.

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Rescue of an influenza A virus wild-type PB2 gene and a mutant derivative bearing a site-specific temperature-sensitive and attenuating mutation.

Content Provider	PubMed Central
Author	Subbarao, E. K. Kawaoka, Y. Murphy, B. R.
Abstract	Live attenuated influenza A virus vaccines are currently produced by the transfer of attenuating genes from a donor virus to new epidemic variants of influenza A virus, with the selection of reassortant viruses that possess the protective antigens (i.e., the two surface glycoproteins) of the epidemic virus and the attenuating genes from the donor virus. The previously studied attenuated donor viruses were produced by conventional methods such as passage of virus at low temperature or chemical mutagenesis. The present paper describes a new strategy for the generation of a donor virus bearing an attenuating, non-surface-glycoprotein gene. This strategy involves the introduction of attenuating mutations into the cDNA copy of the PB2 polymerase gene by site-directed mutagenesis, transfection of in vitro RNA transcripts of PB2 cDNA, and recovery of the transfected PB2 gene into an infectious virus. An avian-human influenza A virus PB2 single-gene reassortant virus (with an avian influenza A virus PB2 gene) that replicates efficiently in avian tissue but poorly in mammalian cells was used as a helper virus to rescue a transfected synthetic RNA derived from a human influenza A virus PB2 gene. The desired human influenza A virus mutant PB2 transfectant was favored in this situation because the avian influenza A virus PB2 gene restricts viral replication in mammalian cells in culture, the system used for rescue, thereby providing strong selection for the virus bearing the human influenza A virus PB2 gene. We validated the feasibility of this approach by rescuing the PB2 gene of the wild-type influenza A/Ann Arbor/6/60 virus and a mutant derivative that had a single amino acid substitution introduced at position 265 by site-directed mutagenesis. Previously, this amino acid substitution had been shown to specify both a temperature-sensitive (ts) and an attenuation (att) phenotype. The rescued mutant 265 PB2 transfectant virus exhibited the ts and att phenotypes, which confirms that these phenotypes were specified by this single amino acid substitution. The transfectant virus was immunogenic and protected hamsters from subsequent challenge with wild-type virus. The cDNA copy of this influenza A/Ann Arbor/6/60 virus mutant 265 PB2 gene will be used as a substrate for the introduction of additional attenuating mutations by site-directed mutagenesis.
Starting Page	7223
File Format	PDF
ISSN	10985514
e-ISSN	10985514
Journal	Journal of Virology
Issue Number	12
Volume Number	67
Language	English
Publisher Date	1993-12-01
Access Restriction	Open
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Virology Immunology Microbiology Insect Science

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