Extracellular signal-regulated activation of Rap1 fails to interfere in Ras effector signalling.
The small GTPase Rap1 has been implicated in both negative and positive control of Ras-mediated signalling events. We have investigated which extracellular signals can activate Rap1 and whether this activation leads to a modulation of Ras effector signalling, i.e. the activation of ERK and the small GTPase Ral. We found that Rap1 is rapidly activated following stimulation of a large variety of growth factor receptors. These receptors include receptor tyrosine kinases for platelet-derived growth factor (PDGF) and epithelial growth factor (EGF), and G protein-coupled receptors for lysophosphatidic acid (LPA), thrombin and endothelin. At least three distinct pathways may transduce a signal towards Rap1 activation: increase in intracellular calcium, release of diacylglycerol and cAMP synthesis. Surprisingly, activation of endogenous Rap1 fails to affect Ras-dependent ERK activation. In addition, we found that although overexpression of active Rap1 is able to activate the Ral pathway, activation of endogenous Rap1 in fibroblasts does not result in Ral activation. Rap1 also does not negatively influence Ras-mediated Ral activation. We conclude that activation of Rap1 is a common event upon growth factor treatment and that the physiological function of Rap1 is likely to be different from modulation of Ras effector signalling.
Laboratory for Physiological Chemistry, Utrecht University, Universiteitsweg 100, 2584 CG Utrecht, The Netherlands.
F J
FJ
Zwartkruis
R M
RM
Wolthuis
N M
NM
Nabben
B
B
Franke
J L
JL
Bos
D016475:3T3 Cells / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000502:physiology*; D008246:Lysophospholipids / Q000494:pharmacology; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D016716:PC12 Cells / Q000187:drug effects / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D011518:Proto-Oncogene Proteins / Q000502:physiology*; D051381:Rats; D015398:Signal Transduction / Q000502:physiology*; D010738:Type C Phospholipases / Q000187:drug effects / Q000502:physiology; D020725:ral GTP-Binding Proteins; D020668:rap GTP-Binding Proteins; D018631:ras Proteins / Q000502:physiology*
D008246:Lysophospholipids; D010982:Platelet-Derived Growth Factor; D011518:Proto-Oncogene Proteins; D004815:Epidermal Growth Factor; D020928:Mitogen-Activated Protein Kinases; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins; D020725:ral GTP-Binding Proteins; D020668:rap GTP-Binding Proteins; D018631:ras Proteins