Molecular characterization of the cellular receptor for poliovirus.
The expression of the human poliovirus receptor (hPVR) in several cultured cell lines was studied with the use of different antibodies directed against hPVR proteins. Immunoprecipitations of metabolically labeled cell lysates revealed that membrane-bound glycoforms of hPVR proteins have a molecular weight of about 80 kDa. By applying inhibitors of the glycosylation pathway (deoxymannojirimycin and swainsonine) we were able to monitor the modification of the hPVR glycoproteins when passing through the processing pathway. We show that a 67-kDa hPVR protein identified earlier (using a vaccinia virus expression system) is an intermediate glycoform probably located in the endoplasmic reticulum or cis-Golgi. Further modification of this glycoform is blocked by vaccinia virus infection or by the inhibitor deoxymannojirimycin. We, therefore, conclude that the 80-kDa glycoforms identified here are the fully processed hPVR isoforms. Surface iodination confirms that only the 80-kDa glycoforms are expressed on the cell surface. Treatment with various deglycosylating enzymes (N-Glycanase, O-Glycanase, Endo-H, and neuraminidase) demonstrates that the hPVR proteins bear sialylated complex-type oligosaccharides. Deglycosylation of the hPVR proteins also reveals the presence of both hPVR membrane-bound forms in cultured cells. Their relative expression levels, with respect to each other, vary considerably. The distribution of these hPVR isoforms in tissues may help explain the natural function of the hPVR proteins.
Bernhardt
G
G
Department of Microbiology, SUNY at Stony Brook 11794-8621.
Bibb
J A
JA
Bradley
J
J
Wimmer
E
E
D000818:Animals; D002460:Cell Line; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D017955:Poliovirus / Q000235:genetics* / Q000378:metabolism; D011233:Precipitin Tests; D011991:Receptors, Virus / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000378:metabolism
D011991:Receptors, Virus; D014759:Viral Envelope Proteins