The short and long forms of the alpha subunit of the stimulatory guanine-nucleotide-binding protein are unequally redistributed during (-)-isoproterenol-mediated desensitization of intact S49 lymphoma cells.
We report here that desensitization of the beta-adrenergic receptor-triggered transmembrane signalling in S49 wild-type lymphoma cells, induced by (-)-isoproterenol (1 microM), results in unequal intracellular redistribution of the splicing variants of the alpha subunit of the stimulatory guanine-nucleotide-binding regulatory (Gs alpha) protein (Gs alpha-short and Gs alpha-long) and alters the functional characteristics of the membrane-associated signal transduction complex. We found that two cellular pools of membranes, light-density membranes and plasma membranes prepared by sucrose-density-gradient centrifugation of cell homogenates differed in their content of Gs alpha splicing subforms and, moreover, that prolonged activation of the beta-adrenergic pathway induced intermembrane redistribution of the splicing variants of Gs alpha. Short (10 min) as well as prolonged (1 h) (-)-isoproterenol treatment of the cells shifted Gs alpha-short from light-density membranes to plasma membranes and increased the total amount of light-density membrane-bound Gs alpha-long; in parallel, the maximal (-)-isoproterenol-stimulated or AlF4(-)-stimulated adenylyl cyclase activities measured in the plasma membrane pools prepared from treated cells decreased. The functional characteristics of the membrane-bound Gs alpha pools were examined by a cyc(-)-reconstitutive adenylyl cyclase assay where extracts of the plasma membrane and light-density-membrane pools, respectively, were mixed with plasma membranes derived from the mutant S49 cell line, cyc-, lacking Gs alpha. The maximal cyc(-)-reconstitutive activities of the extracts prepared from light-density membranes of short-term as well as long-term desensitized cells increased compared to control cells. These findings may indicate differences in the functioning of the splicing variants of Gs alpha.
Wallenberg Laboratory for Cardiovascular Research, Gothenburg University, Sweden.
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D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D000242:Cyclic AMP / Q000096:biosynthesis; D019204:GTP-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D007545:Isoproterenol / Q000494:pharmacology*; D008223:Lymphoma / Q000201:enzymology / Q000378:metabolism*; D051379:Mice; D014407:Tumor Cells, Cultured
D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D007545:Isoproterenol