Proteolytic cleavage of the murine coronavirus surface glycoprotein is not required for its fusion activity.
The surface glycoprotein (S) of the murine hepatitis coronavirus MHV normally undergoes proteolytic cleavage during transport to the cell surface. To determine whether the cleavage of the MHV-JHM S glycoprotein is required to activate its ability to fuse cellular membranes, the protease recognition sequence in a cDNA copy of the S gene was altered from Arg-Arg-Ala-Arg-Arg into Ser-Val-Ser-Gly-Gly by site directed mutagenesis. The mutated and wild type S genes were expressed by means of recombinant vaccinia viruses and it could be shown that the mutated S protein was not cleaved when it was expressed in mouse DBT cells, in contrast to the wild type S protein. Nevertheless, the non-cleaved S protein induced extensive syncytium formation in mouse DBT cells. These results clearly indicate that the non-cleaved form of the MHV S protein is able to mediate cell membrane fusion. Thus, proteolytic cleavage is not an absolute requirement for its fusion function.
Stauber
R
R
Institut für Virologie, Universität Würzburg.
Pfleiderer
M
M
Siddell
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001254:Astrocytoma; D002459:Cell Fusion; D003588:Cytopathogenic Effect, Viral; D010450:Endopeptidases / Q000378:metabolism; D006367:HeLa Cells / Q000382:microbiology; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000235:genetics / Q000502:physiology*; D011499:Protein Processing, Post-Translational; D064370:Spike Glycoprotein, Coronavirus; D014407:Tumor Cells, Cultured / Q000382:microbiology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
D008562:Membrane Glycoproteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus; D010450:Endopeptidases