Processing and routage of HIV glycoproteins by furin to the cell surface

Proteolytic activation of HIV-1 and HIV-2 envelope glycoprotein precursors (gp160 and gp140, respectively) occurs at the carboxyl side of a consensus motif consisting of the highly basic amino acid sequence. We have shown previously (Hallenberger et al., 1997) and confirmed in this report, that furin and PC7 can be considered as the putative physiological enzymes involved in the proteolytic activation of the HIV-1 and HIV-2 envelope precursors. In this study, we show by cell surface biotinylation and immunoprecipitation of the cell surface associated viral glycoproteins with antibodies that the mature viral envelope glycoproteins are correctly transported to the cell membrane. Furthermore, we show that the uncleaved forms of the glycoproteins (gp160HIV-1 and gp140HIV-2) are also highly represented at the cell surface. First, transient expression of gp160 and gp140 into CV1, a cell line known to be inefficient in the proteolytic processing of the env gene, results in the expression of gp160 and gp140 at the cell surface. Moreover, HIV-1 infection of T cells also showed that gp160 is directed to the cell surface. In addition, we show that the precursor is not incorporated in the virus particle following the budding from the cell surface. Furthermore, a gp160 mutant (deficient for three carbohydrate sites on the gp41), shown to be poorly processed with the coexpressed endoproteases, is found to be transported as an uncleaved precursor to the cell surface. In contrast to HIV envelope glycoproteins, the influenza hemagglutinin precursor (HA0), that is thought to be matured by the furin-like enzymes as well, is found to be retained within the cell and is not able to reach the cell surface. Taken together, these results show that the proteolytic maturation of the viral envelope precursors of human immunodeficiency viruses type 1 and type 2 is not a prerequisite for cell surface targeting of the HIV glycoproteins. Implications of these results for antiviral immune response are discussed.