title: Role of Ca2+ influx in bombesin-induced mitogenesis in Swiss 3T3 fibroblasts.
abstract: In Swiss 3T3 fibroblasts a peptide mitogen bombesin, which acts through the phospholipase C-protein kinase C signaling pathway, stimulates DNA synthesis in a manner strictly dependent on the medium calcium concentration: [3H]thymidine incorporation into DNA in the presence of a saturating concentration of bombesin (10(-8) M) is 4-fold greater at 3.0 mM extracellular calcium as compared with a value obtained at 0.03 mM calcium. In the present study we attempted to identify the site and the mechanism of action of Ca2+ influx along the bombesin-induced mitogenic signaling pathway, by comparing bombesin effects at 0.03 and 3.0 mM of medium calcium. Bombesin induces the same extent of increases in [3H]inositol phosphates after 1 min, and comparable sustained increases in the cellular content of 1,2-diacylglycerol for up to 4 h, at either 0.03 or 3.0 mM calcium. Bombesin induces the same extent of phosphorylation of MARCKS protein, the major cellular substrate for protein kinase C, irrespective of the medium calcium concentration for at least 4 h. Moreover, diverse cellular responses elicited by bombesin, including c-fos expression, activation of microtubule-associated protein 2 kinase and S6 kinase, glucose uptake, and protein synthesis but not the release of arachidonic acid and its metabolites, are induced similarly at either 0.03 or 3.0 mM calcium. Down-regulation of cellular protein kinase C nearly completely abolishes bombesin effects on c-fos expression, S6 kinase activation, glucose uptake, and DNA synthesis. These results suggest that the target of Ca2+ influx in bombesin-induced mitogenic signaling pathway is not located along the phospholipase C-protein kinase C signal transduction system including cellular events in early G1 phase that exist downstream to protein kinase C action.
authors.affiliation: Department of Physiology, Faculty of Medicine, University of Tokyo, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Takuwa
authors.forename: A
authors.initials: A
authors.lastname: Iwamoto
authors.forename: M
authors.initials: M
authors.lastname: Kumada
authors.forename: K
authors.initials: K
authors.lastname: Yamashita
authors.forename: Y
authors.initials: Y
authors.lastname: Takuwa
mesh_terms: D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D015152:Blotting, Northern; D001839:Bombesin / Q000494:pharmacology*; D002118:Calcium / Q000502:physiology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002460:Cell Line; D003035:Cobalt / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis; D004075:Diglycerides / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation; D007295:Inositol Phosphates / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008757:Methylglucosides / Q000378:metabolism; D051379:Mice; D008938:Mitosis; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D010750:Phosphoproteins / Q000378:metabolism; D014176:Protein Biosynthesis; D011493:Protein Kinase C / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016760:Proto-Oncogene Proteins c-fos; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction
chemical_list: D001095:Arachidonic Acids; D004075:Diglycerides; D007295:Inositol Phosphates; D047908:Intracellular Signaling Peptides and Proteins; C000619711:Marcks protein, mouse; D008565:Membrane Proteins; D008757:Methylglucosides; D010750:Phosphoproteins; D011506:Proteins; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D012333:RNA, Messenger; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D016718:Arachidonic Acid; D003035:Cobalt; D004247:DNA; D011494:Protein Kinases; D011493:Protein Kinase C; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D001839:Bombesin; D002118:Calcium