- Volume 69, Issue 10, 1988
Volume 69, Issue 10, 1988
- Animal
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Antigenic Relationship between Hantaviruses Analysed by Immunoprecipitation
More LessSummaryAntigenic relationships between seven hantaviruses isolated in Sweden, Belgium, Korea, European U.S.S.R. and Asian U.S.S.R. were studied by radioimmunoprecipitation assays (RIPA) and indirect immunofluorescence tests (IFT). Animal immune sera and haemorrhagic fever with renal syndrome (HFRS) patient sera from the above countries were used. The strains fell into three antigenic groups by both RIPA and IFT:nephropathia epidemica (NE)-type, Korean haemorrhagic fever (KHF)-type and urban rat-type. This antigenic grouping conforms to the clinical grouping of the respective HFRS patients. The major cross-reactive antigen between the antigenic groups was the nucleocapsid protein. Of the two viral glycoproteins, the G2 exhibited weak cross-reactivity between the groups, while the G1 protein appeared to be the least cross-reactive. Patient sera collected at different intervals after the onset of disease showed a marked difference in their capacity to immunoprecipitate the homologous viral glycoproteins, although by IFT all sera had high titres. Host species-related antibody response differences were found by both RIPA and IFT. Using patient sera, a one-way cross-reactivity was seen between the NE-type and the KHF-type viruses. However, animal immune sera clearly demonstrated a reciprocal cross-reactivity between the two virus groups.
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Detection of the Human Rhinovirus Minor Group Receptor on Renaturing Western Blots
More LessSummaryThe human rhinovirus minor group receptor was extracted from HeLa cell membranes and partially purified. Receptor activity was detected on Western blots by binding of 35S-labelled human rhinovirus serotype 2 to the immobilized protein at a position corresponding to an M r of 120 × 103.
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Neutralization of Mouse Xenotropic Virus by Lipoproteins Involves Binding to the Virions
More LessSummaryWe examined the properties of the mouse serum lipoprotein responsible for specific neutralization of the endogenous mouse xenotropic (X-tropic) type C retrovirus. The anti-X-tropic virus activity of the mouse lipoprotein neutralizing factor (NF) could not be blocked by sugars or lectins. Moreover, neutralization did not involve rupture of the virion core. Sucrose density gradient analysis of X-tropic virus mixed with various lipoprotein preparations demonstrated that the binding of complete virions to NF is associated with the neutralization.
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Integration into the Cellular Genome of a Friend Murine Leukaemia Virus Containing a Selectable Marker Reveals a Clonal Origin of Erythroleukaemia
More LessSummaryWe have constructed a selectable Friend murine leukaemia virus (F-MuLV) with a suppressor tRNA (supF) gene integrated into the proviral long terminal repeat. The viral construct was infectious and pathogenic and retained the marker gene when growing in vitro or in vivo. Only a few integration sites of the provirus were detected by Southern blot analysis of the DNA of erythroleukaemic cells. These results indicate that F-MuLV-induced erythroleukaemia is of clonal origin and suggest that insertional mutagenesis is involved in pathogenesis.
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Cooperation in Oncogenesis between BK Virus Early Region Gene and the Activated Human c-Harvey ras Oncogene
More LessSummaryRapidly growing, undifferentiated brain tumours were induced in newborn Syrian hamsters by intracerebral inoculation of a recombinant DNA (pBK/c-rasA) carrying the BK virus (BKV) early region gene and the activated human c-Harvey-ras (c-Ha-ras) oncogene. Neither of the two genes inoculated alone nor recombinant DNA of the BKV early region gene and the normal human c-Ha-ras proto-oncogene were tumourigenic. Tumour-derived cell lines propagated in culture were immortalized and had growth characteristics consistent with a fully transformed phenotype. Tumours and tumour cell lines contained pBK/c-rasA sequences integrated into cellular DNA and expressed BKV- and c-Ha-ras-specific transcripts as well as BKV T antigen and c-Ha-ras p21. These findings are discussed in relation to a possible cooperation or synergism between BKV and cellular oncogenes in human neoplasia.
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Myristylation of Rotavirus Proteins
More LessSummaryAfter labelling with [3H]myristic acid during replication the structural proteins VP6 and VP2 of purified rotavirus particles were found to be myristylated in an amide bond. [3H]Palmitic acid was converted to myristic acid before bonding to the viral proteins. Possible functions of rotavirus protein myristylation are discussed.
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- Plant
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Immunocytochemical Location of Pathogenesis-related b1 Protein Induced in Tobacco Mosaic Virus-infected or Polyacrylic Acid-treated Tobacco Plants
More LessSummaryPathogenesis-related b1 (PR-b1) protein and other serologically related PR-b proteins, known to be associated with both tobacco mosaic virus infection and chemical treatments, such as with polyacrylic acid (PAA), were detected by immunofluorescence microscopy around epidermal, palisade and lacunar leaf cells in hypersensitive tobacco plants (PAA+ line). After gentle fixation and embedding in Lowicryl K4M, PR-b proteins revealed by immunogold labelling were located in the cytoplasm and apoplast. After fixation and embedding in Epon, the PR-b proteins were found to accumulate mainly in the intercellular spaces. These observations support results obtained from biochemical tests.
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Inoculation with RNAs 1 and 2 of Cucumber Mosaic Virus Induces Viral RNA Replicase Activity in Tobacco Mesophyll Protoplasts
More LessSummaryThe activity of membrane-bound RNA-dependent RNA polymerase in tobacco mesophyll protoplasts was measured and in vitro products were characterized after inoculation with cucumber mosaic virus (CMV) or with different combinations of the four virus RNAs and satellite RNA. Activity increased rapidly 4 to 6 h after inoculation with virus particles. When the inoculum contained RNA 1, RNA 2 and RNA 3, most of the in vitro products of the enzyme were full-length positive-sense RNAs (RNAs 1, 2, 3 and 4) that migrated as dsRNA; inoculation of RNA 1 or RNA 2 alone did not increase RNA polymerase activity and no virus RNA was produced. After inoculation with a mixture of RNA 1 and RNA 2, however, the enzyme activity increased and full-length positive-sense RNA 1 and RNA 2 were synthesized, and we speculate that the induced enzyme is CMV RNA replicase as a complex comprising the translation products of RNA 1 and RNA 2, membrane components of the host cell and CMV RNA templates. Protoplasts inoculated with RNA 1, RNA 2 and RNA 4 did not synthesize RNA 4 which suggests that positive-sense RNA 4 does not serve as a template for the replicase complex. Satellite RNA was synthesized in protoplasts inoculated with RNA 1, RNA 2 and satellite RNA which suggests that satellite RNA is replicated by the replicase complex.
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- Fungal
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Molecular Cloning of Hypovirulence-associated Double-stranded RNA in Endothia parasitica and Detection of Sequence-related Single-stranded RNA
More LessSummaryRecombinant plasmids containing cDNA copies of the dsRNAs present in one hypovirulent strain of Endothia (Cryphonectria) parasitica, EP713, were constructed and analysed by restriction endonuclease mapping and Southern hybridization. Overlapping inserts of four plasmids were found to represent most of the large dsRNA (L-dsRNA) sequences. Inserts which represented the two termini of the L-dsRNAs, designated ‘homopolymer’ and ‘heteropolymer’, were identified. These plasmids were used as probes in Northern hybridization experiments in an attempt to detect other RNAs having sequences related to those of the L-dsRNAs. No additional RNAs were detected that hybridized to plasmids representing the middle region of L-dsRNAs. However, plasmids representing the termini of L-dsRNAs hybridized to several RNAs in EP713 ranging in size from 300 to 1300 nucleotides. These RNAs were absent from EP155, the isogenic virulent strain of E. parasitica. RNAs related to the homopolymer terminus were more abundant than those related to the heteropolymer terminus. All were sensitive to digestion by S1 nuclease but resistant to RNase III, indicating that they were single-stranded. Only those ssRNAs related to the homopolymer terminus of L-dsRNAs were retained on oligo(dT)-cellulose. Single-stranded M13 phage DNAs containing the insert of one of the plasmids, in each orientation, were used as probes in Northern hybridization experiments. Hybridization to the ssRNAs was observed with only one of these probes, indicating that the transcripts are derived from only one strand of the L-dsRNAs. These results establish the existence of a set of poly(A)-containing ssRNAs that are 3′-coterminal with the homopolymer terminus of L-dsRNAs and have the same polarity as the poly(A)-containing strand of L-dsRNAs.
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- Addendum
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Structure-Function Analysis of Mouse Interferon Alpha Species: MuIFN-α10, a Subspecies with Low Antiviral Activity
More LessJournal of General Virology (1988), 69, 67–75
It has been drawn to our attention that the DNA sequence of the MuIFN-α10 gene published in this paper is almost identical to one previously published for the MuIFN-α7 gene [Dion, M., Vodjdani, G. & Doly, J. (1986). Sequence and expression of a novel murine interferon alpha gene — homology with enhancer elements in the regulatory region of the gene. Biochemical and Biophysical Research Communications 138, 826–834]. There is one sequence difference in the 5′ non-coding region and one in the 3′ non-coding region, but the protein-coding region is identical in both analyses. It is proposed that MuIFN-α10 be renamed MuIFN-α7(10).
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