Amino-terminal substitutions in the CCR5 coreceptor impair gp120 binding and human immunodeficiency virus type 1 entry

Tatjana Dragic, Alexandra Trkola, Steven W. Lin, Kirsten A. Nagashima, Francis Kajumo, Lu Zhao, William C. Olson, Lijun Wu, Charles R. Mackay, Graham P. Allaway, Thomas P. Sakmar, John P. Moore, Paul J. Maddon

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Abstract

The CC-chemokine receptor CCR5 is required for the efficient fusion of macrophage (M)-tropic human immunodeficiency virus type 1 (HIV-1) strains with the plasma membrane of CD4+ cells and interacts directly with the viral surface glycoprotein gp120. Although receptor chimera studies have provided useful information, the domains of CCR5 that function for HIV-1 entry, including the site of gp120 interaction, have not been unambiguously identified. Here, we use site-directed, alanine-scanning mutagenesis of CCR5 to show that substitutions of the negatively charged aspartic acid residues at positions 2 and 11 (D2A and D11A) and a glutamic acid residue at position 18 (E18A), individually or in combination, impair or abolish CCR5-mediated HIV-1 entry for the ADA and JR-FL M-tropic strains and the DH123 dual-tropic strain. These mutations also impair Env-mediated membrane fusion and the gp120-CCR5 interaction. Of these three residues, only D11 is necessary for CC-chemokine-mediated inhibition of HIV-1 entry, which is, however, also dependent on other extracellular CCR5 residues. Thus, the gp120 and CC- chemokine binding sites on CCR5 are only partially overlapping, and the former site requires negatively charged residues in the amino-terminal CCR5 domain.

Original languageEnglish
Pages (from-to)279-285
Number of pages7
JournalJournal of Virology
Volume72
Issue number1
Publication statusPublished - 1 Jan 1998
Externally publishedYes

Cite this

Dragic, T., Trkola, A., Lin, S. W., Nagashima, K. A., Kajumo, F., Zhao, L., ... Maddon, P. J. (1998). Amino-terminal substitutions in the CCR5 coreceptor impair gp120 binding and human immunodeficiency virus type 1 entry. Journal of Virology, 72(1), 279-285.