Adaptation of HIV-1 to cells with low expression of the CCR5 coreceptor

•HIV-1 can adapt to CD4-positive cells expressing progressively lower levels of CCR5.•The adapted virus' Env changes are located outside of the gp120 CCR5-binding region.•The gp120 of the adapted virus did not increase its affinity for CCR5.•The Env changes enhance HIV-1 replication at all levels of CCR5 expression.•The adapted virus Envs exhibit increased propensity to change conformation.

The binding of the human immunodeficiency virus (HIV-1) envelope glycoprotein (Env) trimer ((gp120/gp41)3) to the receptors CD4 and CCR5 triggers virus entry into host cells. To identify Env regions that respond to CCR5 binding, HIV-1 was serially passaged on a CD4-positive canine cell line expressing progressively lower levels of CCR5. HIV-1 replication was observed in cells expressing ~1300 CCR5 molecules/cell. Env changes that conferred this low-CCR5 replication phenotype were located outside of the known CCR5-binding region of the gp120 Env subunit and did not apparently increase CCR5 binding affinity. The adaptation-associated changes, located in the gp120 α1 helix and in the gp41 HR1 heptad repeat and membrane-proximal external region (MPER), enhanced HIV-1 replication in cells at all levels of CCR5 expression. The adapted Envs exhibited a greater propensity to undergo conformational changes, as evidenced by increased exposure of conserved regions near the CD4- and CCR5-binding sites.