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The tobacco mosaic virus (TMV) 30K movement protein (MP) gene was inserted into a full-length cDNA clone of barley stripe mosaic virus (BSMV) RNAβ replacing the triple gene block (TGB). The resulting recombinant ND-MPT genome, consisting of infectious wt transcripts of BSMV RNAs α and γ, together with the hybrid RNAβ transcript, was inoculated onto test plants to study the functional compatibility...
Thein vitroDNA-binding activity of the C2 protein of tomato yellow leaf curl geminivirus (TYLCV) was studied following its expression inEscherichia colias a fusion protein with an His tag N-terminal extension (His-C2). Southwestern blotting experiments demonstrated that the C2 protein is able to bind both single-stranded and double-stranded DNA probes. In electrophoretic mobility shift assays performed...
Recent reports have highlighted a potential antiviral activity for nitric oxide (NO). The purpose of this study was to investigate the production of NO in mice during vaccinia virus (VV) or herpes simplex virus type 1 infection, and to assess the role of NO in clearance of VV. Reactive nitrogen intermediates (RNI; NO and its stable oxidation products, nitrite and nitrate) were significantly elevated...
To investigate the mechanisms regulating baculovirus virulence and host range we have begun to studyChoristoneura fumiferananuclear polyhedrosis virus (CfMNPV) and its gene expression in permissive and nonpermissive cells. We have identified and mapped three genes on the CfMNPV genome. Thepolyhedringene is located from 0.0 to 2.0 m.u. and two other genes,dnapolandp143,both of which are essential for...
The gene coding for the large subunit of herpes simplex virus type 2 ribonucleotide reductase (RR) (ICP10) has a unique 5′ terminal domain the product of which has a serine/threonine (Ser/Thr) protein kinase (PK) catalytic domain preceded by a transmembrane (TM) segment. Because ICP10 localizes on the cell surface and is internalized by the endocytic pathway like an activated growth factor receptor...
Formation of the glycoprotein gH/gL heterooligomer has important implications for understanding the pathology of human herpesvirus-6 (HHV-6)-associated disease because this complex is essential for infectivity and fusogenic cell-to-cell spread. Definition of the HHV-6 gH domain involved in protein–protein interactions was addressed by targeting regions defined by conserved cysteines identified by...
The hepatitis B virus posttranscriptional regulatory element (PRE), which resides in the 3′ region of the viral S transcripts, is important for the high-level expression of these mRNA species. We have previously shown that the PRE acts intranuclearlyin cisto allow efficient export of intronless transcripts into the cytoplasm, in a manner that is independent of viraltrans-acting factors. We here present...
Two viruses with a novel hemagglutinin (HA), A/duck/Australia/341/83 and A/shearwater/West Australia/2576/79, have been isolated from a duck and a shorebird in Australia. Hemagglutination inhibition and double immunodiffusion assays failed to reveal cross-reactivity with any of the known subtypes (H1 to H14). We therefore propose that these viruses constitute a new HA subtype, H15. Sequence analysis...
Mutant ts10 is an RNA-negative temperature-sensitive mutant of Mahoney type 1 poliovirus. Mutant ts10 3D pol was purified from infected cells and was shown to be rapidly heat-inactivated at 45° when compared to wild-type polymerase. Sequencing of mutant ts10 genomic RNA revealed a U to C transition at nt 7167 resulting in an amino acid change of methionine 394 of 3D pol to threonine...
Rotavirus is one of very few viruses that utilizes the endoplasmic reticulum (ER) for maturation. The maturation process is unique not only because it involves translocation of subviral particles across the ER membrane, but also because mature virus is thought to be retained in the ER until cell lysis. Brefeldin A (BFA) is a compound that blocks protein export from the ER to the Golgi complex and...
Analysis of encapsidated RNA from the Q strain of CMV (Q-CMV) has indicated the presence of a discreet population of molecules of approximately 300 nt termed RNA 5 (Peden, K. W. C., and Symons, R. H.,Virology53, 487–492, 1973.). Q-CMV RNA 5 was isolated and the 5′-end sequence was determined by direct RNA sequencing. This sequence corresponded to the exact beginning of the imperfectly conserved 3′-terminal...
Epstein–Barr virus (EBV) recombinants with theBamHI C promoter (Cp) deleted were compared with wild-type Cp recombinants derived in parallel for their ability to initiate and maintain latent infection and growth transformation in primary human B lymphocytes. Cp-deleted recombinants infected, transformed, and immortalized B lymphocytesin vitroas efficiently as wild-type Cp recombinant EBV. Lymphoblastoid...
It has been suggested that four amino acids which are absolutely conserved in the nsP1 nonstructural proteins encoded by togaviruses and in the homologous proteins encoded by plant viruses in the Sindbis virus (SV) superfamily may constitute a “methyltransferase motif.” In the Sindbis virus nsP1 protein (540 amino acids) these four amino acids are represented by His 39 , Arg 91 , Asp...
The eukaryotic translation initiation factor eIF4E, the cap-binding protein, seems to play an essential role in the establishment of the host shut-off after viral infection. Infection with adenovirus and influenza virus caused dephosphorylation of eIF4E and an involvement of a viral protein was suggested. In this report, we studied several other viruses for their ability to change the phosphorylation...
The IR6 protein of different plaque isolates from three passages of the equine herpesvirus 1 strain Rac was investigated. Southern blot and DNA sequence analyses revealed that plaque isolates from the 12th passage (RacL11 and RacL22) retained both copies of the IR6 gene, whereas two different genotypes were observed by the 185th passage: RacM24 still harbored both copies of the IR6 gene, whereas RacM36...
Previous studies (Mertenset al., Virology157, 375–386, 1987) have shown that removal of the outer capsid layer from bluetongue virus (BTV) significantly reduces (approximately × 10 −4 ) the infectivity of the resultant core particle for mammalian cells (BHK 21 cells). In contrast, the studies reported here, using a cell line (KC cells) derived from a species ofCulicoidesthat can act as a vector...
We have found previously that, in contrast to the free O initiator protein of λ phage or plasmid rapidly degraded by theEscherichia coliClpP/ClpX protease, the λO present in the replication complex (RC) is protected from proteolysis. In amino acid-starvedE. coli relAcells, a temperature shift from 30 to 43° did not affect RC integrity, as judged from the unchanged level of stable λO observed; however,...
The human papillomaviruses associated with cervical cancer (e.g., HPV-16 and HPV-18) express an E7 oncoprotein which mediates the immortalization of primary genital keratinocytes and the transformation of rodent cells. The 105-amino-acid HPV-18 E7 protein contains two zinc fingers as well as a conserved amino-terminal motif (Rb-binding core) which binds and alters the interactions of the retinoblastoma...
High-titer stocks ofrev-defective HIV-1 virions have been produced and characterized. To produce these stocks, CEM cells transduced with a murine retroviral vector that expresses Rev were used to support the growth of an HIV-1 genome containing four inactivating mutations in therevgene. The resulting viral stocks were of high-titer when used to infect CEM cells that expressed Rev, but had no measurable...
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