title: Glycoprotein incorporation and HIV-1 infectivity despite exchange of the gp160 membrane-spanning domain.
abstract: We have examined the role of the membrane-anchoring domain of the HIV-1 glycoproteins in viral glycoprotein function, glycoprotein incorporation, and viral infectivity. For this purpose, we initially exchanged the entire membrane-spanning region with that from a cellular glycoprotein (CD22). Subsequently, the strictly conserved arginine in the central position of the transmembranal alpha-helix was replaced by a neutral residue (R696 --> I696). We have further examined the requirements within the cytoplasmic C-terminus for glycoprotein incorporation and replaced this region of gp160 with the long cytoplasmic C-terminus (118 amino acids) from CD22. Our results show that the specific amino acid sequence of the membrane-spanning region of gp160 is not necessary for viral infectivity, thus making it unlikely that this region is specifically involved in membrane fusion, in glycoprotein incorporation, or in infectivity of the cell lines tested. In contrast, recombinant gp160 with the CD22 C-terminal region, although present at the cell surface and membrane fusion-competent, was excluded from incorporation into particles. This could indicate that steric exclusion, and no pseudotyping, occurs when the heterologous, cytoplasmic C-terminal region is too long and not fitting.
authors.affiliation: Forschungsschwerpunkt Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, Heidelberg, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Wilk
authors.forename: T
authors.initials: T
authors.lastname: Pfeiffer
authors.forename: A
authors.initials: A
authors.lastname: Bukovsky
authors.forename: G
authors.initials: G
authors.lastname: Moldenhauer
authors.forename: V
authors.initials: V
authors.lastname: Bosch
mesh_terms: D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000378:metabolism*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism*; D015815:Cell Adhesion Molecules; D002460:Cell Line; D002462:Cell Membrane / Q000821:virology; D015686:Gene Products, env / Q000378:metabolism*; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D006367:HeLa Cells; D006801:Humans; D037102:Lectins; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000378:metabolism*; D051918:Sialic Acid Binding Ig-like Lectin 2
chemical_list: D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C497867:CD22 protein, human; D015815:Cell Adhesion Molecules; D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D037102:Lectins; D008562:Membrane Glycoproteins; D011498:Protein Precursors; D051918:Sialic Acid Binding Ig-like Lectin 2