title: CCR5-Mediated human immunodeficiency virus entry depends on an amino-terminal gp120-binding site and on the conformational integrity of all four extracellular domains.
abstract: The human immunodeficiency virus type 1 coreceptor activity of CCR5 depends on certain polar and charged residues in its amino-terminal domain. Since studies of chimeric receptors have indicated that the extracellular loops of CCR5 are also involved in viral fusion and entry, we have explored the role of bulky, polar and nonpolar residues in these regions. Selected amino acids in the three extracellular loops were individually changed to alanines, and the coreceptor activities of the mutant CCR5 proteins were tested in a luciferase reporter virus-based entry assay. We found that the cysteines in the extracellular loops of CCR5 are essential for coreceptor activity. However, only minor (two- to threefold) effects on coreceptor function were noted for all of the other alanine substitutions. We also demonstrated that when the first 19 residues of the amino-terminal region were separated from the rest of CCR5, by insertion of glycine/serine spacers between proline 19 and cysteine 20, coreceptor function decreased. Together with our previous studies, these data indicate that both an amino-terminal gp120-binding site and extracellular domain geometry play a role in viral entry.
authors.affiliation: Aaron Diamond AIDS Research Center, The Rockefeller University, New York, New York 10016, USA.
authors.forename: S
authors.initials: S
authors.lastname: Genoud
authors.forename: F
authors.initials: F
authors.lastname: Kajumo
authors.forename: Y
authors.initials: Y
authors.lastname: Guo
authors.forename: D
authors.initials: D
authors.lastname: Thompson
authors.forename: T
authors.initials: T
authors.lastname: Dragic
mesh_terms: D019943:Amino Acid Substitution; D001665:Binding Sites; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D019713:Receptors, CCR5 / Q000737:chemistry* / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D015699:HIV Envelope Protein gp120; D019713:Receptors, CCR5; D003545:Cysteine