title: Variants of human immunodeficiency virus type 1 that efficiently use CCR5 lacking the tyrosine-sulfated amino terminus have adaptive mutations in gp120, including loss of a functional N-glycan.
abstract: By selecting the R5 human immunodeficiency virus type 1 (HIV-1) strain JR-CSF for efficient use of a CCR5 coreceptor with a badly damaged amino terminus [i.e., CCR5(Y14N)], we previously isolated variants that weakly utilize CCR5(Delta18), a low-affinity mutant lacking the normal tyrosine sulfate-containing amino-terminal region of the coreceptor. These previously isolated HIV-1(JR-CSF) variants contained adaptive mutations situated exclusively in the V3 loop of their gp120 envelope glycoproteins. We now have weaned the virus from all dependency on the CCR5 amino terminus by performing additional selections with HeLa-CD4 cells that express only a low concentration of CCR5(Delta18). The adapted variants had additional mutations in their V3 loops, as well as one in the V2 stem (S193N) and four alternative mutations in the V4 loop that eliminated the same N-linked oligosaccharide from position N403. Assays using pseudotyped viruses suggested that these new gp120 mutations all made strong contributions to use of CCR5(Delta18) by accelerating a rate-limiting CCR5-dependent conformational change in gp41 rather than by increasing viral affinity for this damaged coreceptor. Consistent with this interpretation, loss of the V4 N-glycan at position N403 also enhanced HIV-1 use of a different low-affinity CCR5 coreceptor with a mutation in extracellular loop 2 (ECL2) [i.e., CCR5(G163R)], whereas the double mutant CCR5(Delta18,G163R) was inactive. We conclude that loss of the N-glycan at position N403 helps to convert the HIV-1 envelope into a hair-trigger form that no longer requires strong interactions with both the CCR5 amino terminus and ECL2 but efficiently uses either site alone. These results demonstrate a novel functional role for a gp120 N-linked oligosaccharide and a high degree of adaptability in coreceptor usage by HIV-1.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, OR 97239-3098, USA.
authors.forename: Emily J
authors.initials: EJ
authors.lastname: Platt
authors.forename: Danielle M
authors.initials: DM
authors.lastname: Shea
authors.forename: Patrick P
authors.initials: PP
authors.lastname: Rose
authors.forename: David
authors.initials: D
authors.lastname: Kabat
mesh_terms: D000220:Adaptation, Biological; D000577:Amides / Q000494:pharmacology; D019943:Amino Acid Substitution; D002460:Cell Line; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D000077560:Enfuvirtide; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000494:pharmacology / Q000502:physiology; D023581:HIV Fusion Inhibitors / Q000494:pharmacology; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D009154:Mutation; D010446:Peptide Fragments / Q000235:genetics / Q000494:pharmacology / Q000502:physiology; D011134:Polysaccharides / Q000737:chemistry / Q000502:physiology; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D019713:Receptors, CCR5 / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication
chemical_list: D000577:Amides; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D023581:HIV Fusion Inhibitors; C059762:HIV envelope protein gp120 (305-321); D010446:Peptide Fragments; D011134:Polysaccharides; D000644:Quaternary Ammonium Compounds; D019713:Receptors, CCR5; D000077560:Enfuvirtide; C119369:TAK 779