Receptors induce chemotaxis by releasing the βγ subunit of G i , not by activating G q or G s

Abstract

Many chemoattractants cause chemotaxis of leukocytes by stimulating a structurally distinct class of G protein-coupled receptors. To identify receptor functions required for chemotaxis, we studied chemotaxis in HEK293 cells transfected with receptors for nonchemokine ligands or for interleukin 8 (IL-8), a classical chemokine. In gradients of the appropriate agonist, three nonchemokine G i -coupled receptors (the D 2 dopamine receptor and opioid μ and δ receptors) mediated chemotaxis; the β 2 -adrenoreceptor and the M3-muscarinic receptor, which couple respectively to G s and G q , did not mediate chemotaxis. A mutation deleting 31 C-terminal amino acids from the IL-8 receptor type B quantitatively impaired chemotaxis and agonist-induced receptor internalization, but not inhibition of adenylyl cyclase or stimulation of mitogen-activated protein kinase. To probe the possible relation between receptor internalization and chemotaxis, we used two agonists of the μ-opioid receptor. Morphine and etorphine elicited quantitatively similar chemotaxis, but only etorphine induced receptor internalization. Overexpression of two βγ sequestering proteins (βARK-ct and α t ) prevented IL-8 receptor type B-mediated chemotaxis but did not affect inhibition of adenylyl cyclase by IL-8. We conclude that: ( i ) Nonchemokine G i -coupled receptors can mediate chemotaxis. ( ii ) G i activation is necessary but probably not sufficient for chemotaxis. ( iii ) Chemotaxis does not require receptor internalization. ( iv ) Chemotaxis requires the release of free βγ subunits.