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Volume 65,
Issue 11,
1984
Volume 65, Issue 11, 1984
- Animal
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Interaction between Epstein–Barr Virus-determined Nuclear Antigen (EBNA) and the Viral DNA
More LessSUMMARYEpstein–Barr virus (EBV) nuclear antigen (EBNA) was purified from the Burkitt lymphoma line Raji and its EBV DNA-binding properties were characterized. EBNA binding protected fragments of about 30 bp of B95-8 cell-derived EBV DNA from an excess of DNase I. Human anti-EBNA antibodies prevented DNA binding. Purified extracts from EBNA-negative cells did not protect EBV DNA against DNase I digestion. Mapping of the EBV DNA fragments protected from endonuclease (EcoRI, HindIII, SalI) digestion revealed many binding sites. Similar results were obtained following mixing of crude cell extracts and HindIII-digested fragments of EBV DNA and subsequent immunoprecipitation of the EBNA-DNA complex. In experiments involving the analysis of EBV DNA, fragments were protected from DNase I digestion by purified EBNA.
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Recovery of Host Cell Protein Synthesis during Infection with a Temperature-sensitive Mutant of Newcastle Disease Virus
More LessSUMMARYWhen chick embryo fibroblasts infected with high multiplicities of a non-revertible temperature-sensitive (ts) mutant (ts53) of Newcastle disease virus (NDV) at the permissive temperature (34 °C) and incubated to allow the appearance of virus-induced proteins were shifted up to and incubated at the non-permissive temperature (42 °C), analysis of [3H]leucine-labelled proteins by SDS-PAGE revealed a marked increase in cellular protein synthesis and a decline in viral synthesis, with an eventual return to an apparently uninfected state. Mutant ts53 apparently ceased to suppress cellular protein synthesis, and translation of viral proteins was markedly inhibited. This was not observed in ts +-infected cells, or in ts53-infected cells maintained at the permissive temperature. Recovery from the infected state was inhibited by actinomycin D at any stage, but was not prevented by cycloheximide present immediately before and during temperature shift-up. Mutant ts53 was shown to be RNA− at 42 °C, although viral mRNAs synthesized at 34 °C are shown to be present throughout host recovery but in an inactive state.
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Cell Division Does Not Affect Sendai Virus Genome Replication in Persistently Infected BHK Cells
More LessSUMMARYThe extent of Sendai virus genome replication in persistently infected BHK cells actively growing or at confluence was followed by estimation of the [3H]uridine incorporated into intracellular nucleocapsid RNA. First, we showed that, in the presence of actinomycin D, actively growing persistently infected cells were taking up threefold more [3H]uridine than resting cells. This higher uptake exhibited by growing cells was observed neither in persistently infected cells in the absence of actinomycin D, nor in acutely infected cells in the presence of actinomycin D. Assuming that the cellular pool of unlabelled uridine stays constant, we used a correction factor for this difference in [3H]uridine uptake and estimated [3H]uridine incorporation in nucleocapsid RNA, normalizing the data either to the amount of cell or of viral template. Results showed that the viral genome replication, expressed either way, was not significantly influenced by cell growth conditions.
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- Fungal
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Infection of Protoplasts of the Wheat Take-all Fungus, Gaeumannomyces graminis var. tritici, with Double-stranded RNA Viruses
More LessSUMMARYProtoplasts from a virus-free isolate, 3b1a conD/A3, of Gaeumannomyces graminis var. tritici were inoculated with a purified preparation of a mixture of four viruses, A, B1, B2 and C, from G. graminis isolate 3b1a in the presence of polyethylene glycol. Three out of 30 fungal colonies obtained by regeneration of inoculated protoplasts were found to be infected, two with virus B1 and one with virus B2. The concentrations of these viruses in the newly infected isolates were similar to their respective concentrations in isolate 3b1a and remained stable through three serial subcultures.
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