Temperature-dependent internalization of virus glycoproteins in cells infected with a mutant of Semliki Forest virus.
When the ts-1 mutant of Semliki Forest virus (SFV) was grown in chick embryo or BHK 21 cells at the restrictive temperature (39 degrees C), its membrane glycoproteins were arrested in the endoplasmic reticulum, but started to migrate to the cell surface once the cultures were shifted to the permissive temperature (28 degrees C). If the temperature of infected cells was raised back to 39 degrees C, ts-1 glycoproteins disappeared from the cell surface as evidenced by loss of surface immunofluorescence and by radioimmunoassay based on the binding of 125I-labeled protein A. This phenomenon was specific for ts-1 at 39 degrees C as it was observed neither in cells infected with wild-type SFV at 39 degrees C nor with ts-1 at 28 degrees C. The disappearance of the ts-1 glycoproteins was due to internalization. The internalized proteins were digested, as shown by specific decrease of virus glycoproteins labelled with [35S]methionine at 39 degrees C before shift to 28 degrees C, and by concomitant release of acid soluble 35S-activity into the culture medium. Ts-1 infected cells were treated before shift back to 39 degrees C with Fab' fragments, prepared from IgG against the viral membrane glycoproteins. After shift back to 39 degrees C, the Fab' fragments disappeared from the cell surface. In the presence of chloroquine, they could be visualized in vesicular structures, using an anti-IgG-fluorescein isothiocyanate conjugate. The internalization of ts-1 glycoproteins was not inhibited by carbonylcyanide p-trifluoromethoxy phenylhydrazone, chloroquine, cytochalasin B, vinblastine, colcemid, or monensin.
D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002738:Chloroquine / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D007700:Kinetics; D008985:Monensin / Q000494:pharmacology; D009154:Mutation; D012672:Semliki forest virus / Q000235:genetics*; D013696:Temperature; D014764:Viral Proteins / Q000378:metabolism*
D007140:Immunoglobulin Fab Fragments; D014764:Viral Proteins; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D002738:Chloroquine; D008985:Monensin