Vaccinia virus-mediated expression and identification of the human poliovirus receptor.
We have used cDNA encoding the cellular receptor for poliovirus (PVR) to prepare polyclonal antisera against beta-galactosidase PVR fusion proteins. One of these antisera allowed identification of a glycoprotein doublet band of about 67 kDa in membrane preparations of HeLa cells and in a PVR cosmid-bearing mouse cell line. In vitro translation of PVR-specific transcripts gave rise to a protein of 46 kDa; the product had a molecular weight of 67 kDa when microsomal membranes were added to the cell-free extract. Overexpression of PVR cDNA in mouse L-cells by means of a recombinant vaccinia virus led to the synthesis of a glycoprotein having a molecular weight identical to that of the glycosylated in vitro product. The vaccinia virus-mediated protein was also recognized by a monoclonal antibody that blocks poliovirus infection. Its biological activity was demonstrated by poliovirus binding and infectivity assays. The data show that PVR is a glycoprotein of 67 kDa and that this protein is sufficient to confer poliovirus susceptibility to mouse cells.
Zibert
A
A
Department of Microbiology, State University of New York, Stony Brook 11794.
Selinka
H C
HC
Elroy-Stein
O
O
Moss
B
B
Wimmer
E
E
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins; D051379:Mice; D008970:Molecular Weight; D017955:Poliovirus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011233:Precipitin Tests; D011991:Receptors, Virus / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics
D006023:Glycoproteins; D008565:Membrane Proteins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; C066100:poliovirus receptor; D004247:DNA