Human immunodeficiency virus type 1 infection of SK-N-MC cells: domains of gp120 involved in entry into a CD4-negative, galactosyl ceramide/3' sulfo-galactosyl ceramide-positive cell line

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Published inJournal of Virology Vol. 69; no. 12; pp. 7383 - 7390
Main Authors Harouse, J M, Collman, R G, González-Scarano, F
Format Journal Article
LanguageEnglish
Published United States American Society for Microbiology 01.12.1995
Subjects
AIDS/HIV
Antigens, CD - genetics
Antigens, CD - physiology
Base Sequence
Binding Sites
CD4 Antigens - genetics
CD4 Antigens - physiology
Cell Line
Chromatography, High Pressure Liquid
DNA Primers
DNA, Viral - analysis
DNA, Viral - isolation & purification
Enzyme-Linked Immunosorbent Assay
Galactosylceramides - analysis
Galactosylceramides - physiology
HeLa Cells
HIV Envelope Protein gp120 - metabolism
HIV-1 - genetics
HIV-1 - physiology
human immunodeficiency virus
Humans
Kinetics
Molecular Sequence Data
Neuroblastoma
Phenotype
Polymerase Chain Reaction
Receptors, Virus - analysis
Receptors, Virus - physiology
Species Specificity
Sulfoglycosphingolipids
Tumor Cells, Cultured
Online AccessGet full text

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Abstract Article Usage Stats Services JVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue JVI About JVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0022-538X Online ISSN: 1098-5514 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JVI .asm.org, visit: JVI       
AbstractList The primary receptor for human immunodeficiency virus (HIV) is the CD4 molecule; however, in vitro evidence suggests that a neutral glycolipid, galactosyl ceramide (GalCer) or a derivative molecule, 3' sulfogalactosyl ceramide (GalS), may serve as an alternative receptor for HIV type 1 (HIV-1) in cells of neural and colonic origin. Biochemical studies have demonstrated that recombinant gp120 envelope protein binds to GalCer/GalS in both solid-phase enzyme-linked immunosorbent assay and high-performance thin-layer chromatography overlays. We have used the SK-N-MC cell line, a CD4-negative, GalCer/GalS-positive cell line previously characterized as susceptible to HIV-1 infection, to identify virus isolates with either a positive infection phenotype, HIVHxB2, or a negative infection phenotype, HIV-1(89.6). Using a solid-phase virus binding assay, we determined the level of restriction in HIV-1(89.6) infection to be at the level of virus-glycolipid binding. Furthermore, using HIV-1HxB2-HIV-1(89.6) chimeras, we have identified a 193-amino-acid fragment from the envelope region of HIV-1HxB2 containing the V3, V4, and V5 regions which confers a positive infection phenotype on the HIV-1(89.6) background. Recombinant viruses which separate this 193-amino-acid fragment into two distinct chimeras are each able to confer a positive infection phenotype on the background of HIV89.6, suggesting that a stable GalCer/GalS-envelope interaction is dependent on the conformation of the envelope protein in the context of the viral membrane. Alternatively, the GalCer/GalS-gp120 bond may involve multiple sites on the oligomeric envelope protein.
Article Usage Stats Services JVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue JVI About JVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0022-538X Online ISSN: 1098-5514 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JVI .asm.org, visit: JVI       
The primary receptor for human immunodeficiency virus (HIV) is the CD4 molecule; however, in vitro evidence suggests that a neutral glycolipid, galactosyl ceramide (GalCer) or a derivative molecule, 3' sulfo-galactosyl ceramide (GalS), may serve as an alternative receptor for HIV type 1 (HIV-1) in cells of neural and colonic origin. Biochemical studies have demonstrated that recombinant gp120 envelope protein binds to GalCer/GalS in both solid-phase enzyme-linked immunosorbent assay and high-performance thin-layer chromatography overlays. We have used the SK-N-MC cell line, a CD4-negative, GalCer/GalS-positive cell line previously characterized as susceptible to HIV-1 infection, to identify virus isolates with either a positive infection phenotype, HIV sub(HxB2), or a negative infection phenotype, HIV-1 sub(89.6). Using a solid-phase virus binding assay, we determined the level of restriction in HIV-1 sub(89.6) infection to be at the level of virus-glycolipid binding. Furthermore, using HIV-1 sub(HxB2)-HIV-1 sub(89.6) chimeras, we have identified a 193-amino-acid fragment from the envelope region of HIV-1 sub(HxB2) containing the V3, V4, and V5 regions which confers a positive infection phenotype on the HIV-1 sub(89.6) background. Recombinant viruses which separate this 193-amino-acid fragment into two distinct chimeras are each able to confer a positive infection phenotype on the background of HIV sub(89.6), suggesting that a stable GalCer/GalS-envelope interaction is dependent on the conformation of the envelope protein in the context of the viral membrane. Alternatively, the GalCer/GalS-gp120 bond may involve multiple sites on the oligomeric envelope protein.
Author R G Collman
F González-Scarano
J M Harouse
AuthorAffiliation Department of Neurology, School of Medicine University of Pennsylvania, Philadelphia 19104-6146, USA
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Snippet Article Usage Stats Services JVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley...
The primary receptor for human immunodeficiency virus (HIV) is the CD4 molecule; however, in vitro evidence suggests that a neutral glycolipid, galactosyl...
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StartPage 7383
SubjectTerms AIDS/HIV
Antigens, CD - genetics
Antigens, CD - physiology
Base Sequence
Binding Sites
CD4 Antigens - genetics
CD4 Antigens - physiology
Cell Line
Chromatography, High Pressure Liquid
DNA Primers
DNA, Viral - analysis
DNA, Viral - isolation & purification
Enzyme-Linked Immunosorbent Assay
Galactosylceramides - analysis
Galactosylceramides - physiology
HeLa Cells
HIV Envelope Protein gp120 - metabolism
HIV-1 - genetics
HIV-1 - physiology
human immunodeficiency virus
Humans
Kinetics
Molecular Sequence Data
Neuroblastoma
Phenotype
Polymerase Chain Reaction
Receptors, Virus - analysis
Receptors, Virus - physiology
Species Specificity
Sulfoglycosphingolipids
Tumor Cells, Cultured
Title Human immunodeficiency virus type 1 infection of SK-N-MC cells: domains of gp120 involved in entry into a CD4-negative, galactosyl ceramide/3' sulfo-galactosyl ceramide-positive cell line
URI http://jvi.asm.org/content/69/12/7383.abstract
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