Virology
During serial undiluted passage of rubella virus (RUB) in Vero cells, two species of defective-interfering (DI) RNAs of approximately 7000 and 800 nucleotides (nts) in length were generated (Frey, T. K., and Hemphill, M. L., Virology 164, 22–29, 1988). In this study, these DI RNAs were characterized by molecular cloning, hybridization with probes of defined sequence, and primer extension. The 7000-nt...
Attempts to define the genetic determinants required for efficient growth of human immunodeficiency virus type 1 (HIV-1)in monocyte-macrophages were made by constructing chimeras between two infectious clones of HIV-1 (HXB2 and LW/C), which despite only minor differences in their DNA sequence have striking differences in cell tropism. Although both of them replicate efficiently in peripheral blood...
Purified M protein of VSV has been reported to aggregate at low NaCl concentration. Using light scattering, analyticalcentrifugation, and electron microscopy (EM), we have studied this phenomenon. Our results demonstrate that self aggregation of M protein can be reversed by increasing the salt concentration. Below 250 mM NaCl, there is an equilibrium between aggregates and monomeric M protein. Most...
We used a spleen necrosis virus-based retroviral vector to introduce the polyomavirus replication origin into rat cells and developed a system to analyze homologous recombination events that do not reconstitute a selectable marker. Introduction of the gene coding for the polyomavirus large T antigen into the cell lines by DNA transfection promoted high-frequency recombination between the two retroviral...
The life cycle of human papillomaviruses (HPVs) is dependent on epithelial differentiation. Among the viral proteinsexpressed in differentiated epithelial cells are the viral capsid proteins, L1 and L2, as well as the E1^E4 fusion proteins. In this study, the expression and intracellular localization of the E1^E4 proteins of HPV type 31b were examined in both monolayer and raft cultures of the CIN-612...
We isolated two recombinant baculoviruses each of which expresses a varicella-zoster virus (VZV) homolog of one ofthe seven herpes simplex virus type 1 (HSV-1) genes required for DNA replication. We performed transient origin-dependent DNA replication assays in insect cells in which we substituted a baculovirus which expresses a VZV protein for a baculovirus which expresses its HSV homolog. VZV gene...
Cellular ezrin-radixin-moesin (ERM) family proteins, members of the actin-binding proteins of the band 4.1 superfamily, were detected in the virions of enveloped viruses, such as rabies, vesicular stomatitis, Newcastle disease, and influenza viruses. To elucidate the mechanism of ERM protein incorporation, we investigated possible association of ERM proteins with viral components in rabies virus-infected...
A set of 150 synthetic peptides spanning the proteins NS3-NS4-NS5 of the hepatitis C virus (HCV) was synthesizedand tested with a panel of 20 sera obtained from HCV-infected patients. Of 62 peptides prepared from the NS3 region, none exhibited strong antigenic reactivity. Rather, five peptides from this region demonstrated specific reactivity with only 5–10% of anti-HCV-positive sera. Nonetheless,...
HERV-H elements are a large family of endogenous retrovirus-like sequences found in approximately 1000 dispersedcopies in the genomes of humans and other primates. The most abundant subclass of these elements is a partially deleted form of 5.8 kb which is transcribed primarily as a 5.6-kb unit length RNA and a 3.7-kb spliced derivative. The provirus-like structure of these elements suggests that their...
A full-length cDNA clone of rice yellow mottle sobemovirus (RYMV) was synthesized and placed adjacent to a bacteriophageT7 RNA polymerase promoter sequence. Capped-RNA transcripts produced in vitro were infectious when mechanically inoculated onto rice plants (Oryza sativa L.). Individual full-length clones varied in their degree of infectivity but all were less infectious than native viral RNA. A...
The RNA-binding activity of the interferon-inducible, RNA-dependent protein kinase PKR, expressed from the human PKRcDNA, was quantitated using a gel mobility-shift assay. The N-terminal R-domain truncation Wt(1–243) and the full-length catalytic mutant K296R(1–551) were analyzed for their abilities to bind adenovirus VA, RNA, human immunodeficiency virus TAR RNA, and the synthetic homopolymer pl:pC...
The role of the 385 nucleotide 5′ noncoding region (NCR) in the translation of the pestivirus genome was investigated. In vitro translation of an RNA transcript containing the 5′ NCR of the bovine viral diarrhea virus (BVDV) genome followed by the coding sequence of the first gene product (p20) of the BVDV large open reading frame resulted in the synthesis of a 20-kDa polypeptide. Results from hybrid-arrest...
Cellular expression of the herpes simplex virus type 1 thymidine kinase (HSV1-TK) gene promotes cell death in the presence of specific nucleoside analog substrates such as acyclovir (ACV). We have reported that lymphoid CD4+ cells harboring an HSV1-TK gene, under the transcriptional control of the HIV-1 long terminal repeat (HUT-TK), are completely protected from HIV-1 spread in the presence of 10...
Borna disease is a chronic neurological disease caused by an enveloped negative-strand RNA virus (BDV). Experimentaldisease can be reproduced in rats with brain homogenates derived from infected animals or with virus derived from infected cells in culture. The virus replicates in cultured cells without evidence of cytopathic effect or production of significant levels of cell-free virus. Borna disease...
Previous studies have found Kunjin (KUN) virus isolates from within Australia to be genetically homogenous and that the envelope protein of the type strain (MRM61C) was unglycosylated and lacked a potential glycosylation site. We investigated the extent of antigenic variation between KUN virus isolates from Australia and Sarawak using an immunoperoxidase assay and a panel of six monoclonal antibodies...
Since the pathogenesis of SIV mac disease complex is thought to be explained by the tropism of the infecting virus for either CD4 + T-lymphocytes or macrophages or both types of cells, we compared the infection in primary macaque macrophages with molecularly cloned, lymphocyte-tropic SIV mac 239 and a cloned, macrophage-tropic chimeric virus (SIV mac 239/17E) whose...
Previously, we reported two cDNAs derived from the Marek's disease virus (MDV) long internal repeat region. A 14-kDapolypeptide (p14) encoded by two small open reading frames (ORFs) from at least two distinct cDNAs is expressed in cells lytically infected with both oncogenic and attenuated MDV as well as in cells latently infected and transformed by MDV. In this study, we demonstrate that p14 is serotype...
The tegument of bovine herpesvirus-1 (BHV-1) carries an abundant protein of 96 kDa, termed VP8. Immunolabeling usingVP8-specific antiserum and colloidal gold-labeled protein A as the electron-dense marker was used to identify VP8 in the virions and virus-infected cells. VP8 was confirmed to be a tegument protein that, like the herpes simplex virus-1 homologue VP13/14, contains O-linked carbohydrates...
A murine model of genital infection with a thymidine kinase-deficient (tk − ) strain of herpes simplex virus type 2 (HSV-2) was utilized to examine the local urogenital antibody response to HSV-2. Vaginal inoculation with HSV-2 tk − protected against a subsequent genital challenge with a lethal dose of virulent HSV-2. After primary vaginal infection, predominantly HSV-specific IgG...