Different effects of a single amino acid substitution on three adjacent epitopes in the gp41 C-terminal tail of a neutralizing antibody escape mutant of human immunodeficiency virus type 1.
The envelope protein of human immunodeficiency virus type 1 (HIV-1) comprises the outer gp 120 SU domain and the anchoring gp41 TM domain, and the conventional view is that it has a single transmembrane region with the following C-terminal sequence situated entirely within the virion. However, we have recently proposed that the gp41 C-terminal region comprises three transmembrane regions and an external loop structure. Part of this loop is the peptide 731PRGPDRPEGIEEEGGERDRDRS752 that carries three antibody epitopes, 734PDRPEG739, 740IEEE743, and 746ERDRD750. PDRPEG is not detected in virions but reacts with its cognate MAb (C8) in Western blots, IEEE is a linear and non-neutralizing epitope, and ERDRD is a conformational and neutralizing epitope. Here we show that escape mutants selected with neutralizing ERDRD-specific antibody had a single 732R-->G substitution, 14 residues upstream of the cognate epitope, and no longer bound the selecting antibody. The same amino acid substitution altered epitope PDRPEG in the virion so that it now reacted with MAb C8, but left epitope IEEE unaffected. Introduction of 732R-->G by site-specific mutagenesis into the gp41 of cloned HIV-1 NL4-3 virions allowed them to escape neutralization by ERDRD-specific IgG, and confirms that 732R makes a major contribution to the neutralizing conformation of the 731-752 region of the C-terminal tail of gp41.
Department of Biological Sciences, University of Warwick, Coventry, UK.
L
L
McLain
J L
JL
Brown
L
L
Cheung
S A
SA
Reading
C
C
Parry
T D
TD
Jones
S M
SM
Cleveland
N J
NJ
Dimmock
D019943:Amino Acid Substitution; D000596:Amino Acids / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes / Q000235:genetics; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D006801:Humans; D046911:Macromolecular Substances; D016297:Mutagenesis, Site-Directed; D009500:Neutralization Tests; D010455:Peptides / Q000276:immunology; D011485:Protein Binding
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D015700:HIV Envelope Protein gp41; D046911:Macromolecular Substances; D010455:Peptides