Zx. Wang et al., CCR5 HIV-1 coreceptor activity - Role of cooperativity between residues inN-terminal extracellular and intracellular domains, J BIOL CHEM, 274(40), 1999, pp. 28413-28419
Human (H-) CCR5 is the primary coreceptor for ENV-mediated fusion by R5 str
ains of human immunodeficiency virus type 1, whereas mouse (M-) CCR5 lacks
this function. An array of 23 H/M-CCR5 hybrids containing increasing amount
s of H-CCR5 extending from the N terminus generated by random chimeragenesi
s had a biphasic pattern of coreceptor activity with JRFL and 89.6, reveali
ng active regions in the N-terminal extracellular domain (N-ED) and at the
junction of cytoplasmic loop 3. The M-CCR5 mutant in which divergent residu
es were replaced with the corresponding H-CCR5 N-ED sequence (NyYTsE) gaine
d coreceptor function in fusion but not infection experiments. A M-CCR5 dou
ble mutant with substitution of human sequences for divergent residues from
the N-ED and cytoplasmic loop 3 had augmented coreceptor activity in fusio
n assays and gain of function in infection experiments. The SIV-251 ENV uti
lized H- and M-CCR5 and variants, Flow cytometric analysis of M-CCR5 mutant
s and bifunctional receptors composed of CD4 domains fused to M-CCR5 mutant
s excluded the possibility that differences in coreceptor activity resulted
from variations in cell surface expression. These results demonstrate that
the coreceptor activity of the H-CCR5 N-ED is modulated by intracellular r
esidues, illustrating the complexity of CCR5 requirements for interaction w
ith ENV.