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At least 865 records · Page 48Linked to original sources
PubMed1982

Activation of herpes simplex virus (HSV) type 1 genome by temperature-sensitive mutants of HSV type 2.

Previous studies have demonstrated that herpes simplex viruses (HSV) type 1 (HSV-1) and type 2 (HSV-2) can be maintained in a repressed form in human embryo lung cells. Reducing the incubation temperature or superinfecting with a heterologous herpesvirus, human cytomegalovirus (HCMV), results in activation of virus replication. We now report that superinfection with a partially homologous herpesvirus, HSV-2, also resulted in activation of HSV-1. To minimize excessive synthesis of infectious HSV-2 while allowing virus gene expression, repressed HSV-l-infected cultures were superinfected with HSV-2 temperature-sensitive (ts) mutants (tsF3, tsB5, or tsH9). The predominant virus replicated after HSV-2 ts mutant superinfection at a nonpermissive temperature was identified as activated parental-like HSV-1 by (i) plaquing efficiency at permissive (34 degrees) and nonpermissive (40.5 degrees) temperatures, (ii) sensitivity to inhibition by the HSV-l-specific antiviral agent (E)-5-(2-bromovinyl)-2'-deoxyuridine, and (iii) restriction endonuclease cleavage analysis. In addition, the fact that superinfection with HSV-2 tsB5 or tsH9, which are unable to synthesize virus DNA and express only early virus genes at nonpermissive temperature, resulted in synthesis of virus demonstrated that HSV-2 DNA synthesis is not required for activation. This system has provided the basis for further studies concerning the regulation of HSV gene expression in human cells.

Bromodeoxyuridine↗
PubMed1983

Inhibition of measles virus budding by phenothiazines.

HeLa cells infected with measles virus show an accumulation of virus-specific strands at the plasma membrane after addition of the anticalmodulin drugs trifluoperazine (TFP) and chlorpromazine (CPZ), whereas spherical virus particles are almost completely absent. At low drug concentrations (10-15 microM TFP; 30-40 microM CPZ) the inhibitory effect is dependent on the presence of extracellular calcium. The strands complete the budding process after removal of the drugs. Restoration of virus budding is not sensitive to cycloheximide and immunoprecipitation experiments give evidence that the viral protein synthesis is not qualitatively altered in the presence of TFP. The data indicate that both drugs arrest the budding process at an intermediate stage at the plasma membrane. The inability of the strands to comigrate with cytochalasin B-induced actin patches suggests that the inhibition of budding is probably the result of an impaired interaction of viral structures with the cytoskeleton.

Antipsychotic Agents↗
PubMed1983

Maintenance of human cytomegalovirus genome in human diploid fibroblast cells.

Human embryo lung cells pretreated with a combination of human leukocyte interferon and acyclovir were infected with human cytomegalovirus (HCMV) and treated daily for 14 days with the same inhibitor combination. After removal of the inhibitors, the incubation temperature was maintained at either 37 or 40.5 degrees. Incubation of infected cells at 37 degrees after inhibitor removal resulted in progressive virus-specific cytopathology and eventually total destruction of the cell culture. However, HCMV-infected cells incubated at 40.5 degrees after inhibitor removal exhibited little or no virus-induced cytopathology and HCMV remained noninfectious for 8 days. After extended incubation at 40.5 degrees, infectious HCMV was readily detectable even though virus-specific cytopathology was not evident. Reducing the incubation temperature from 40.5 to 37 degrees resulted in stimulation of infectious virus replication and subsequent destruction of the infected cell culture monolayer within 10 days.

Acyclovir↗
PubMed1983

Identification and characterization of deoxyribonucleoprotein complexes containing the major DNA-binding protein of herpes simplex virus type 1.

Nuclear extracts were prepared from cells infected with herpes simplex virus type 1 (HSV-1) and fractionated by sucrose gradient centrifugation to identify deoxyribonucleoprotein complexes involved in viral replication. Large amounts of an HSV-1 induced protein with a molecular weight of about 133,000 sedimented as a broad peak in the 25 S region of the gradient and cosedimented with 13 S DNA fragments. The sedimentation of both the protein and DNA decreased upon treatment of nuclear extracts with DNase. This result indicated that the protein and DNA were associated in deoxyribonucleoprotein complexes. The protein was identified as the HSV-encoded major DNA-binding protein ICP8 based on its molecular weight, its association with DNA in nuclear extracts, and its immunoprecipitation with monospecific antiserum and monoclonal antibody to ICP8. Deoxyribonucleoprotein complexes containing ICP8 could be immunoprecipitated from nuclear extracts. When DNA was extracted from these immunoprecipitates, fractionated by agarose gel electrophoresis, transferred to nitrocellulose paper, and hybridized to 32P-labeled HSV-1 or cell DNA, both HSV-1 and cell DNA sequences were identified. Cesium chloride gradient analysis of the immunoprecipitated DNA indicated that duplex DNA was present in the complexes. Thus, the major DNA-binding protein of HSV-1 is associated with both duplex HSV-1 and cell DNA in vivo.

DNA-Binding Proteins↗
PubMed1983

Multiple origins of the complementary defective genomes of RF and origin proximal sequences of GS, two human papovavirus isolates.

It has previously been shown that the genome of RF virus, a variant of the human papovavirus, BK, consists of two DNA species, one (R1a) with a deletion corresponding to the early and the other (R2) with a deletion corresponding to the late region of BKV (A. Pater, M.M. Pater, and G. di Mayorca (1980). J. Virol. 36, 480-487; A. Pater, M. M. Pater, R. M. Dougherty, and G. di Mayorca (1981a). Virology 113, 86-94). In this report transfection experiments are used to show that these DNA species are individually defective for infection and that both DNA molecules are required simultaneously for the infection of human embryonic kidney (HEK) cells. DNA fragments containing the origin of replication in each of the DNA species are analyzed to show that R1a contains three and R2 contains two origins of replication. In addition, several changes in the repeat region proximal to the origin of replication are observed. The changes involve deletions and insertions. Examination of the deleted junctions most often reveals involvement of short stretches of repeated sequences (hot spots) in recombination. Another observed change is the insertion into R2 of a 63-bp sequence which contains no homology to either BK or SV40 DNA. This insertion is into the late-promoter region of this late-region coding DNA and appears to replace a poor "TATA" box in BK wild type with a better TATA box with the correct spacing from the "CAAT" box. A 304-bp fragment containing the origin of replication, early and late promoters, and the repeat units proximal to the origin of replication of GS, another variant of human BKV papovaviruses has also been sequenced. Several rearrangements, including deletions and insertions in the repeat region, are observed. Moreover, when homologous regions of this virus are compared to that of BKV, five base changes are detected, one of which is in the 23-bp origin region. This base change gives this 23-bp palindrome of GS a perfect two-fold rotational symmetry.

Cell Line↗
PubMed1987

Generation of competent virus in the REV helper cell line C3.

We have studied the generation of replication-competent virus in cultures of the helper cell line C3, which harbors a packaging-defective provirus derived from reticuloendotheliosis virus. We transfected the C3 line with a defective provirus encoding the chicken growth hormone gene and carrying the HSV-1 tk gene. The appearance of competent virus was assayed by infection of Spafas C/E CEF, monitoring reverse transcriptase activity, and rescue of the TKTU assayed on BRLtk- cells. Our results indicate that cultures of C3 producing TKTU can release viruses which have variable growth characteristics and which can remain latent in culture for extended periods of time.

Animals↗