The glycosylation of human immunodeficiency virus type 1 transmembrane glycoprotein (gp41) is important for the efficient intracellular transport of the envelope precursor gp160.
The role of the glycans of the human immunodeficiency virus type 1 transmembrane glycoprotein (gp41) in the intracellular events of Env precursor (gp160) biosynthesis has been examined by the use of a mutant gp160 in which the cluster of conserved glycosylation sites within the gp41 domain (Asn-621, -630 and -642) has been mutated. Expression of the wild-type and mutant forms of gp160 in BHK-21 cells using recombinant vaccinia viruses has shown that the kinetics of the events occurring in the endoplasmic reticulum (ER) were normal: both Env proteins had similar kinetics of disulphide bond formation, as determined by the acquisition of CD4-binding capability, and both had similar kinetics of oligomer formation. However, in contrast to the parental molecule, mutated gp160 displayed relatively slow transport from the cis to the medial Golgi where it was retained in the oligomeric state. Transport to the trans Golgi was impaired, as determined by the sensitivity of gp160 to glycosidases. Cleavage of mutated gp160 at the gp120/gp41 junction was substantially reduced but this was apparently not due to the involvement of the gp41 glycosylation in the cleavage reaction by furin inasmuch as, in the baculovirus system, mutated gp160 could be cleaved when recombinant furin was co-expressed. The reduced cleavage in mammalian cells may thus reflect the impaired routing of mutated Env to the compartment where cleavage occurs. The glycan component of gp41 is, therefore, important for the efficient intracellular transport and processing of gp160. gp160 lacking gp41 carbohydrates is an additional example, among few others, of a protein lacking glycans that is arrested in the Golgi rather than the ER following its biosynthesis.
Fenouillet
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Institute of Virology, NERC, Oxford, UK.
Jones
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D000595:Amino Acid Sequence; D000818:Animals; D001216:Asparagine; D015704:CD4 Antigens / Q000096:biosynthesis / Q000378:metabolism; D002460:Cell Line; D017124:Conserved Sequence; D006224:Cricetinae; D045683:Furin; D015686:Gene Products, env / Q000096:biosynthesis / Q000378:metabolism*; D005822:Genetic Vectors; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41 / Q000096:biosynthesis / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D007668:Kidney; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D011498:Protein Precursors / Q000096:biosynthesis / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013381:Subtilisins / Q000096:biosynthesis; D014162:Transfection; D014616:Vaccinia virus
D015704:CD4 Antigens; D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D011498:Protein Precursors; D011994:Recombinant Proteins; D001216:Asparagine; D013381:Subtilisins; D045683:Furin