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Rotaviruses are the leading cause of severe diarrhea in infants and young children worldwide. Thus, the development of an effective rotavirus vaccine is a major public health goal. This study was performed to identify the gene or genes responsible for rotavirus virulence or host range restriction and attenuation in a natural host. Such knowledge could have an important bearing on the selection of...
The Epstein-Barr virus (EBV) BZLF1 gene is expressed early upon induction of the viral lytic cycle and its protein product is unique in its ability to disrupt viral latency in some latently infected cell lines. Anti-immunoglobulin (anti-Ig) treatment of the Burkitt's lymphoma cell line Akata, which bears surface IgG, has previously been shown to synchronously induce transcription of the BZLF1 gene...
Infection with the adenovirus type 2 ts1 mutant at the nonpermissive temperature resulted in the production of noninfectious virions. This has been ascribed to the P137L mutation in the virus-encoded cysteine protease which causes a defect in protease activity. Here we have examined the ts1 defect in detail as a means of learning more about the viral protease. The ts1 protease accumulated in the nucleus...
After the unexpected appearance of lethal symptoms on tomato plants infected with the PSTVd strain Intermediate Di, viroids were isolated and sequenced. It was found that a new strain, named RG 1, had been generated spontaneously in our greenhouse. In a different series of plant passages two new strains, named QF A and QF B, were detected which coexisted with the wild-type strain Di. Strains QF A...
The role of N-linked glycosylation in the assembly, intracellular transport, and fusion activity of the paramyxovirus SV5 fusion (F) protein was examined. Each of the six potential glycosylation sites in the F protein was individually removed by oligonucleotide-directed mutagenesis on a cDNA clone encoding the SV5 F protein. When the mutant F proteins were expressed in eukaryotic cells using the vaccinia...
After intranasal instillation of mice with vesicular stomatitis virus (VSV), olfactory receptor neurons are infected. By 12 to 24 hr postinfection, VSV antigens are observed in adjoining supporting and basal cells and in other structures of the olfactory epithelium and lamina propria. Peripheral deafferentation of the olfactory epithelium with Triton X-100 or bilateral surgical bulbectomy does not...
To study paramyxovirus-mediated cell fusion it would be advantageous to express in a cell a single protein that could cause regulated syncytium formation at neutral pH following a specific activation signal. We have constructed two SV5 fusion (F) protein mutants that contain three arginine residues in the cleavage site and two separate glycine to alanine changes in the fusion peptide. The mutants...
Plants mechanically inoculated with soil-borne wheat mosaic virus (SBWMV, Oklahoma isolate) generated several deleted forms of RNA2, the smaller genomic RNA (3593 nt). Four naturally deleted forms were cloned by RT-PCR methods and the sequences around each deletion site were compared SBWMV RNA2 molecules were deleted for 519, 759, 964, or 1030 nt, respectively, each within the coat protein-readthrough...
Herpes simplex virus type 1 immediate-early polypeptide Vmw110 (ICPO) is a general transactivator of gene expression in transfection assays and is required for the fully efficient onset of viral lytic replication. It has also been implicated in the process of viral reactivation from latency. Its mechanism of action is unknown, but any involvement in latency requires interactions between viral and...
Homotypic and heterotypic neutralization determinants of bluetongue virus serotype 17 (BTV-17) were investigated with a panel of five neutralizing monoclonal antibodies (MAbs). One MAb (MAb 034) was originally raised to BTV serotype 10 (BTV-10) but also neutralizes BTV-17 (P. V. Rossitto, and N. J. MacLachlan, 1992, J. Gen. Virol. 73, 1947-1952). Competitive binding studies indicate that the MAbs...
The temperature-sensitive mutant tsB821 of Autographa californica nuclear polyhedrosis virus has two nucleotide transitions in the transregulatory ie 1 gene (Riblero et al. (1994) J. Virol. 68, 1075-1084). These mutations result in two conservative amino acid substitutions in the encoded protein. To determine whether the mutations affect the ability of IE1 to interact with viral enhancer elements,...
Pseudorabies virus (PRV) early protein 0 (EP0) is the homolog of varicella-zoster virus (VZV) open reading frame 61 (ORF61) protein and herpes simplex virus type 1 (HSV-1) ICP0. A PRV EP0 deletion mutant grows poorly in cell culture, suggesting that EP0 plays a critical role in the viral replicative cycle. In this study, we have shown that the growth defect of an EP0 deletion mutant was complemented...
Tobacco plants were transformed with the cucumber mosaic cucumovirus (CMV) 3a gene and the in planta-expressed 3a protein was detected immunologically. The 3a protein was predominantly localized in a subcellular fraction corresponding to the cytosol. Two frameshift and four deletion mutants were created within the 3a open reading frame of CMV RNA 3. Five of these mutants, containing an N-terminal,...
Two DNA constructs encoding portions of the human immunodeficiency virus type-1 (HIV-1) genome have been used to raise antibody responses in BABL/c mice. One DNA (pNL4-3.env) expresses the natural form of the envelope glycoprotein (Env) of HIV-1-NL4-3 (NL4-3). The second (pNL4-3.dpol) produces noninfectious NL4-3 particles. These two DNAs (alone or in combination) raised only transient titers of anti-Env...
Double-subgenomic Sindbis virus (dsSIN) recombinants that express cassettes encoding prM-E or a C-terminally truncated form of E of Japanese encephalitis virus (JEV) were constructed. The products were efficiently expressed in both mammalian and mosquito cell lines infected with the dsSIN recombinants. However, suppression of prM-E secretion from mammalian cells infected with dsSIN-prM-E recombinants...
The herpes simplex virus transactivator VP16 directs the assembly of a multicomponent protein-DNA complex with cellular components Oct-1 and VCAF-1, contributing a potent carboxyl-terminal acidic activation domain that is essential for activation of gene expression in mammalian cells. We show here that VP16, devoid of this acidic activation domain, functions as a strong transcriptional activator in...
Rotavirus variants resistant to neutralization were selected using monoclonal antibodies (N-MAbs) raised to VP7 of rotavirus G types 2, 3, and 6. Their neutralization resistance patterns and deduced VP7 amino acid sequences were obtained. Variants selected by two G2-specific N-MAbs from the homologous parent virus RV-5 showed single amino acid (aa) mutations in the antigenic A region. However, variants...
The immediate-early IE1 protein of Autographa californica nuclear polyhedrosis virus (AcMNPV) is an important regulator of viral gene transcription. To provide a tool for further analysis of the expression and function of IE1, a polyclonal antiserum was raised against IE1 expressed in bacteria. Immunoblot analysis of infected cell lysates was used to monitor the accumulation of IE1 throughout the...
The murine cytomegalovirus α (immediate-early) gene product, IE2 391aa , a protein that is related to the human cytomegalovirus US22 protein family, had previously been shown to be dispensable for viral growth in cell culture. In transient assays, however, this protein was found to transactivate the murine CMV ie1/ie3 and ie2 promoters, as well as a number of other promoters...
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