Deletion of a single N-linked glycosylation site from the transmembrane envelope protein of human immunodeficiency virus type 1 stops cleavage and transport of gp160 preventing env-mediated fusion.
The transmembrane envelope glycoprotein (gp41) of human immunodeficiency virus type 1 possesses four consensus sites (Asn-X-Ser/Thr) for the incorporation of N-linked sugars situated on the extracellular domain of the molecule. The purpose of this investigation was to determine the significance of each of these sites in relation to the structure and function of the viral envelope glycoprotein. Each of the four sites was removed by in vitro mutagenesis of gp160 sequence in the non-infectious viral clone pEVd1443, so that amino acids 616, 621, 642 and 679 were each changed from asparagine to serine. The effects of mutagenesis were assessed by syncytium assay after wild-type or mutant envelope clones had been transfected into CD4+ HeLa cells. Removal of the glycosylation site at position 642 resulted in the synthesis of precursor gp160 that was neither cleaved, to give gp120 and gp41, nor transported to the plasma membrane of transfected cells. A consequence of these events was that envelope mutant 642 failed to induce syncytia between neighbouring cells in which it had been expressed. The results of this study indicate that N-linked glycosylation of Asn-642 in the glycoprotein produced by the pEVd1443 expression system is necessary for the correct intracellular processing of gp160 to yield surface-expressed, fusogenic gp41.
Dash
B
B
Division of Virology, National Institute for Medical Research, Mill Hill, London, U.K.
McIntosh
A
A
Barrett
W
W
Daniels
R
R
D001483:Base Sequence; D001692:Biological Transport; D017931:DNA Primers / Q000737:chemistry; D015967:Gene Expression Regulation, Viral; D015686:Gene Products, env / Q000502:physiology*; D006031:Glycosylation; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41 / Q000502:physiology; D015497:HIV-1 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008561:Membrane Fusion; D008562:Membrane Glycoproteins / Q000502:physiology*; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000502:physiology*; D011499:Protein Processing, Post-Translational
D017931:DNA Primers; D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D008562:Membrane Glycoproteins; D011498:Protein Precursors