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  • 1
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Radioactive-labelled virions and nucleocapsids of rat cytomegalovirus (RCMV) were purified from the supernatant and subcellular fractions of infected rat embryo fibroblasts (REF) and analyzed by SDS-polyacrylamide gel electrophoresis. Nuclear nucleocapsids contain one major protein of 138 kD, which is considered to be the basic invariant structural element of RCMV. Enveloped virions consist of 28 protein species, five of which were clearly identified as glycoproteins (58, 64, 76, 112 and 118 kD). Using pulse labelling procedures on RCMV-infected REF cells, after removal of a previously established translation block by cycloheximide, two RCMV-induced immediate-early (IE) protein species (71 and 85 kD) were both detected in the nuclear and cytoplasmic cell fractions. When pulse-labelling was performed for 16 hours in presence of Actinomycin D, only the 85 kD IE protein was detected in the nucleus. The results indicate that the 85 kD IE polypeptide is of importance in early transcriptional events of viral genes. Protein metabolism studies revealed that late in the course of RCMV infection (at 3 days p.i.) protein synthesis has been dramatically changed. Some cellular proteins are greatly suppressed while other cellular proteins are clearly enhanced. Moreover, active synthesis of 8 new cytoplasmic proteins and 9 new nuclear proteins occurs. Most of these proteins were identified as structural constituents of virions.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Rat cytomegalovirus (RCMV) induces a cytosol thymidine kinase (TK) in G0-phase rat embryo fibroblasts (REF), but not in a TK deficient rat cell line (R-2), though virus titers in both cell types reached comparable levels. The results indicate that TK is neither virus-coded nor is required for a productive infection in R-2 cells. A deoxycytidine kinase (dCK) is induced in either growing or RCMV-infected REF and R-2 cells, suggesting that dCK is essential for both host-cell and viral DNA synthesis. A deoxy-guanosine kinase (dGK) is detectable in low concentrations in either growing or G0-phase REF and R-2 cells suggesting that this enzyme is cell-cycle independent. In contrast, RCMV induces high persisting levels of dGK, particularly in R-2 cells, indicating that this enzyme is of crucial importance for viral DNA synthesis. By comparison of thermostabilities and electrophoretic mobilities (Rf for TK, dCK and dGK were 0.12; 0.97; and 0.54, respectively) the enzymes were found to be substrate specific but of cellular origin. In contrast to TK and dCK, only dGK is inhibited by Acyclovir (Ki=320 µm). It is suggested that RCMV inducable dGK is an important enzyme determining thein vitro anti-CMV activity of Acyclovir.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In this paper the development of a battery of approximately 70 mouse monoclonal antibodies (McAbs) to RCMV-induced antigens and their characterization is discussed. Their reactivity with the whole scala of ca. 30 virus specific proteins was tested in an enzyme linked immunoassay (ELISA) whereas their ability to detect RCMV-antigens at different locations ofin vitro infected cell cultures and at different stages of infection was tested by immunofluorescence. In order to determine to what specific (viral) protein each of these McAbs is directed against we used an immunoprecipitation technique, followed by SDS-PAGE. Furthermore, neutralizing capacity of each McAb was tested, as well as the immunoglobulin class they belong to. In this manner we defined six categories of monoclonal antibodies on the basis of immunofluorescence aspect. The six categories identify most important viral structural proteins.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In 8 of 10 wild rats trapped in The Netherlands, an infectious viruslike agent was isolated predominantly from the salivary glands and could be serially passed in laboratory rats. In rat embryo cells a typical cytomegalo-like cytopathic effect was produced. The morphologic and cultural characteristics of the isolated agent were comparable with those of the mouse cytomegalovirus (MCMV). The virus-nucleocapsid had a size of 92 nm and was not ether-resistant. The extracellular nucleocapsids were often enclosed by an outer layer of very variable shape and size. The formation of Fc receptors on cells infected with the rat virus could be demonstrated. The wild rats possessed neutralizing antibodies to the isolated agent. The rat agent grew only in rat embryo fibroblast cells while MCMV grew in rat and mouse embryo cells. The rat agent gave plaques in REF monolayers. Electron microscope studies showed the presence of nucleocapsids in the nucleus.
    Type of Medium: Electronic Resource
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