Expression of membrane-associated and secreted variants of gp160 of human immunodeficiency virus type 1 in vitro and in continuous cell lines.
The gene encoding the 856-amino-acid envelope glycoprotein, gp160, of human immunodeficiency virus type 1 was mutagenized and transfected into Chinese hamster ovary cells. Continuous cell lines that constitutively produced gp160 variants were isolated and used to study the biosynthesis and orientation of gp160 in cellular membranes. In vivo studies of gp160 variants failed to reveal domains upstream of amino acid residue 665 that could serve as stop transfer sequences. Analyses of gp160 variants expressed in vitro in a translation-coupled translocation system were consistent with the in vivo studies and provided evidence that gp160 is a simple bitopic membrane protein. A model for the orientation and function of gp160 in cellular membranes is presented. The cell lines described provide a convenient source of the gp120-gp41 complex.
Berman
P W
PW
Department of Molecular Biology, Genetech, Inc., South San Francisco, California 94080.
Nunes
W M
WM
Haffar
O K
OK
D000386:AIDS-Related Complex / Q000276:immunology; D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology; D001345:Autoradiography; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D006678:HIV / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D006801:Humans; D007118:Immunoassay; D009154:Mutation; D014176:Protein Biosynthesis; D012191:Retroviridae Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014158:Transcription, Genetic; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins