Attenuation of murine coronavirus infection by ammonium chloride
Identifieur interne : 003497 ( Main/Exploration ); précédent : 003496; suivant : 003498Attenuation of murine coronavirus infection by ammonium chloride
Auteurs : Lee Mizzen [Canada] ; Anne Hilton [Canada] ; Steve Cheley [Canada] ; Robert Anderson [Canada]Source :
- Virology [ 0042-6822 ] ; 1985.
English descriptors
- Teeft :
- Ammonium, Ammonium chloride, Approx, Assay, Cell extracts, Cell fusion, Cheley, Chloride, Coronavirus, Coronavirus replication, Culture medium, Early events, External virus, Extracellular, Extracellular virus, Glycoprotein, Helenius, Herpes simplex virus type, Infectious center assay, Infectivity, Inhibitory, Inhibitory effect, Inoculated, Inoculum, Internalization, Intracellular, Lysosome, Lysosomotropic, Membrane fusion, Mhvinfected cells, Mizzen, Mouse cells, Mouse hepatitis virus, Murine, Normal medium, Polypeptide, Poole, Progeny, Replication, Rna, Semliki, Semliki forest virus, Uncoating, Vesicle, Vesicular stomatitis virus, Viral, Viral envelope, Viral genome, Viral polypeptides, Viral protein synthesis, Virus, Virus inoculum, Virus internalization, Virus progeny, Weak bases.
Abstract
Abstract: Ammonium chloride at a concentration of 20 mM delayed by 4–5 hr the production of virus progeny in mouse L-2 cells infected at high multiplicity with mouse hepatitis virus (MHV). This delay was seen in the production of both intracellular and extracellular virus. However, the final titers were similar to those produced by MHV-infected cells maintained in normal medium. The manifestation of virus-induced cell fusion was similarly found to be delayed, but not otherwise decreased in severity, when ammonium chloride was present in the culture medium. Ammonium chloride caused similar delays in production of virus-specific, positive-sense RNAs and of viral polypeptides. The relative proportions and apparent molecular weights of viral RNAs and polypeptides were similar to those found in MHV-infected cells cultured in normal medium. In vitro translation of endogenously produced viral RNAs in cell extracts, prepared from MHV-infected cells, was not inhibited by ammonium chloride. Thus, ammonium chloride has no specific, inhibitory effect on viral protein synthesis. Ammonium chloride did not reduce the number of virus-infected cells in culture, as monitored by infectious center assay. Analysis of early events in MHV infection showed that ammonium chloride did not affect adsorption or internalization of MHV by L-2 cells. However, the subsequent eclipse phase, as monitored by decline in infectivity of internalized virus inoculum proceeded less efficiently in the presence of ammonium chloride. On the basis of the known inhibitory effects of ammonium chloride on lysosomal/endosomal functions, the results suggest an endosomal mechanism of MHV uncoating. Thus the primary effect of ammonium chloride on MHV infection of L-2 cells is to attenuate virus uncoating, thereby chronologically displacing all subsequent virus-encoded functions.
Url:
DOI: 10.1016/0042-6822(85)90345-9
Affiliations:
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Le document en format XML
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<term>Cheley</term>
<term>Chloride</term>
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<term>Coronavirus replication</term>
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<term>Early events</term>
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<term>Extracellular virus</term>
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<term>Infectious center assay</term>
<term>Infectivity</term>
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<term>Inhibitory effect</term>
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<term>Inoculum</term>
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<term>Membrane fusion</term>
<term>Mhvinfected cells</term>
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<term>Rna</term>
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<term>Semliki forest virus</term>
<term>Uncoating</term>
<term>Vesicle</term>
<term>Vesicular stomatitis virus</term>
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<term>Viral envelope</term>
<term>Viral genome</term>
<term>Viral polypeptides</term>
<term>Viral protein synthesis</term>
<term>Virus</term>
<term>Virus inoculum</term>
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<front><div type="abstract" xml:lang="en">Abstract: Ammonium chloride at a concentration of 20 mM delayed by 4–5 hr the production of virus progeny in mouse L-2 cells infected at high multiplicity with mouse hepatitis virus (MHV). This delay was seen in the production of both intracellular and extracellular virus. However, the final titers were similar to those produced by MHV-infected cells maintained in normal medium. The manifestation of virus-induced cell fusion was similarly found to be delayed, but not otherwise decreased in severity, when ammonium chloride was present in the culture medium. Ammonium chloride caused similar delays in production of virus-specific, positive-sense RNAs and of viral polypeptides. The relative proportions and apparent molecular weights of viral RNAs and polypeptides were similar to those found in MHV-infected cells cultured in normal medium. In vitro translation of endogenously produced viral RNAs in cell extracts, prepared from MHV-infected cells, was not inhibited by ammonium chloride. Thus, ammonium chloride has no specific, inhibitory effect on viral protein synthesis. Ammonium chloride did not reduce the number of virus-infected cells in culture, as monitored by infectious center assay. Analysis of early events in MHV infection showed that ammonium chloride did not affect adsorption or internalization of MHV by L-2 cells. However, the subsequent eclipse phase, as monitored by decline in infectivity of internalized virus inoculum proceeded less efficiently in the presence of ammonium chloride. On the basis of the known inhibitory effects of ammonium chloride on lysosomal/endosomal functions, the results suggest an endosomal mechanism of MHV uncoating. Thus the primary effect of ammonium chloride on MHV infection of L-2 cells is to attenuate virus uncoating, thereby chronologically displacing all subsequent virus-encoded functions.</div>
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