- Volume 67, Issue 4, 1986
Volume 67, Issue 4, 1986
- Animal
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Evidence that the Major Delayed-Early DNA-binding Proteins of Herpesvirus Saimiri Are Bound to DNA in vivo
More LessSummaryAssociations of herpesvirus saimiri-specified proteins with nuclear fractions from cultures of infected cells were probed by nuclease digestion, detergent extractions and immunofluorescence microscopy using monoclonal antibodies to virus polypeptides. Nuclease digestion selectively released delayed-early polypeptides with apparent mol. wt. of 110000 (110K) and 51000 (51K) from nuclei of infected cultures and the majority of each of these polypeptides partitioned with the insoluble fraction after detergent extraction of such nuclei. However, the nuclease-mediated release of both these proteins was specifically reduced when nuclei were isolated from cultures in which virus DNA synthesis had been inhibited with phosphonoacetic acid (PAA). In addition, the 110K polypeptide partitioned into the soluble fraction when nuclei from PAA-treated cultures were extracted with detergent. Immunofluorescence microscopy revealed characteristic and distinctive subnuclear localizations of the 110K and 51K polypeptides in control cultures and these patterns of subnuclear accumulations were markedly altered in cultures treated with PAA. We conclude that the DNA-binding properties of the delayed-early 110K and 51K proteins of herpesvirus saimiri previously observed in vitro are likely to reflect their functions as DNA-binding proteins in vivo.
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Strain-dependent Virulence Characteristics of Bluetongue Virus Serotype 11
More LessSummaryTwo strains of bluetongue virus (BTV) serotype 11, UC-2 and UC-8, were identified by the electrophoretic migration pattern of their genomic RNA segments on polyacrylamide gel electrophoresis. Significant differences in virulence of these two viruses could be demonstrated by subcutaneous inoculation of newborn mice. No signs of disease were observed in mice infected with UC-2. Mice infected with UC-8 died of a severe necrotizing encephalitis, which resembled lesions in bovine and ovine foetuses infected with BTV.
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Cloning and Analysis of Integrated Hepatitis B Virus DNA of the adr Subtype Derived from a Human Primary Liver Cell Carcinoma
More LessSummaryA 10 kb genomic DNA fragment derived from a human primary liver cell carcinoma (PLC) and containing integrated hepatitis B virus (HBV) DNA was cloned and analysed. Physical mapping showed the viral DNA to comprise a linear sequence of at least 2.8 kb (87%) of the HBV genome and to be of the adr subtype. Integration appeared to have occurred in the region of the viral genome spanning the cohesive ends. The cellular flanking DNA sequences to one side of the integrated viral DNA contained repeats of the Alu family. The finding of no apparent rearrangements of the integrated HBV DNA sequences in this clone is in contrast to the situation in the huSP and PLC/PRF/5 PLC cell lines in which the integrated viral DNA sequences are greatly rearranged and suggests that such rearrangements may be atypical of solid PLCs.
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Lack of Detectable Reverse Transcriptase Activity in Human and Chimpanzee Sera with a High Infectivity for Non-A, Non-B Hepatitis
SummaryA serum sample from a patient with hepatitis and samples from two experimentally infected chimpanzees, all with a high infectivity for non-A, non-B hepatitis, were tested for reverse transcriptase. Biopsy confirmed that the hepatocytes of the chimpanzees that received these sera contained the characteristic tubular structures associated with non-A, non-B hepatitis. None of these three sera revealed detectable enzyme activity. We have not been able to confirm the association of reverse transcriptase activity with non-A, non-B hepatitis reported recently.
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Restriction Enzyme Analysis of Granulosis Viruses Isolated from Artogeia rapae and Pieris brassicae
More LessSummaryThirteen isolates of granulosis virus from Artogeia (= Pieris) rapae and two from Pieris brassicae were compared by restriction enzyme analysis. All the isolates gave very similar fragment profiles with XhoI, SmaI and BglI. but at least 11 of them could be distinguished using EcoRI, BstI and HindIII. Similarities and differences between profiles suggested that the isolates could be placed in three subtypes. This subtyping correlated closely with the geographical origin of the isolates, which came from Europe, North America, Asia and Australasia. All the isolates were highly infectious for A. rapae with median lethal dose values for third instar larvae ranging from 102.3 to 102.6. Only two of four isolates of one subtype had significant infectivity for third instar P. brassicae; thus, this broader host range did not correlate with grouping by restriction enzyme analysis.
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Polyoma Virus Middle T Gene Can Trigger Malignant Transformation of Early Passage Rodent Cells
More LessSummaryPrevious studies on the tumourigenic conversion of early passage rat embryo cells by the polyoma virus early genes have suggested a multigenic control of tumourigenesis. Thus, the large T gene can immortalize early passage rat cells and can relieve the serum dependence of normal and transformed cells. The middle T gene alone cannot immortalize early passage cells; however, it can induce cells of established cell lines to become anchorage-independent and tumourigenic. Here we show that when linked to transcriptional enhancers, the polyoma virus middle T gene can trigger the complete malignant transformation of early passage rodent cells. Therefore, the polyoma virus middle T gene does not require a cooperating oncogene to induce malignant conversion of these cells.
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Mode of Sensitivity and Resistance of Vaccinia Virus Replication to Interferon
More LessSummaryIn this study we show that vaccinia virus replication can be sensitive or resistant to interferon (IFN) in the same strain of mouse L cells. When IFN-treated L cells were maintained in suspension culture, infection led to a rapid inhibition of both viral and cellular protein synthesis together with breakdown of viral RNA and of rRNA. When IFN-treated L cells were maintained in monolayer culture, infection did not lead to significant inhibition of viral protein or RNA synthesis and breakdown of viral or of rRNA was not observed. The resistance of vaccinia virus replication to IFN was not dependent on the input multiplicity or state of growth of the cells (actively dividing or resting). Qualitative and quantitative differences in viral transcription and translation were observed between the two virus-cell systems. Our findings are consistent with the hypothesis that the sensitivity or resistance of vaccinia virus to IFN is mediated by specific viral products that act as activators or selective inhibitors of, at least, the dsRNA-dependent ppp(A2′p)nA synthetase/RNase system.
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HVJ (Sendai Virus) Stimulates Release of Interferon from Leukocytes Used Once for Interferon Production
More LessSummaryLeukocytes, subjected once to interferon (IFN) induction by HVJ (Sendai virus), were studied for their capability to produce IFN after a second similar stimulus. Substantial amounts of IFN (about 30000 IU/ml) were recovered. Experiments using cycloheximide or actinomycin D and kinetic studies showed that this IFN originated mainly in IFN which resided within the cell as a result of the first induction and was released after the second stimulation. Increasing amounts of HVJ used for the second stimulus resulted in proportionally increased yields of IFN, reaching a plateau at the same dose of HVJ (1000 HAU/ml) as that which gave optimal yields after the first stimulation. Evidence is presented that the capacity of HVJ to trigger the production of a second IFN harvest is closely associated with its infectivity.
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The Use of a Monoclonal Antibody Specific for the N-terminal Region of Southern Bean Mosaic Virus as a Probe of Virus Structure
More LessSummaryThe N-terminal cyanogen bromide peptide of the coat protein of the cowpea strain of southern bean mosaic virus (SBMV-C) was purified by high-pressure liquid chromatography and used as an immunogen in the production of monoclonal antibodies. One monoclonal antibody, designated 4D6, bound to both purified peptide and immobilized virus in a solid-phase ELISA. The reactivity of this monoclonal antibody with immobilized peptide could not be inhibited by native SBMV-C but was inhibited by low levels of EDTA-swollen virus in an antigen inhibition ELISA. The binding of 4D6 to the swollen conformation of the virus was not significantly diminished upon collapsing the swollen virus by pH adjustment or addition of calcium ions. Reactivity of the antibody was also observed with native virus particles which had been dialysed against weakly alkaline buffers. The binding of 4D6 was extremely sensitive to trypsin proteolysis of the virus coat protein, indicating that the antibody binding site was most likely located within the first 30 amino acid residues of the N-terminus. Monoclonal antibody 4D6 also cross-reacted weakly with red clover necrotic mosaic virus in both indirect ELISA and antigen inhibition ELISA.
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