Phenylarsine oxide stimulates a cytosolic tyrosine kinase activity and glucose transport in mouse fibroblasts.
In the present report we further approach the mechanism by which insulin and phenylarsine oxide (PAO), a trivalent arsenical compound, regulate glucose transport in mouse fibroblasts (NIH3T3). First, we show that PAO is a powerful stimulatory agent on glucose transport. Second, at least three series of observations indicate that this action of PAO is not mediated through the insulin receptor: (i) the same effect of PAO is observed in NIH3T3 and in transfected cells expressing 6 x 10(6) insulin receptors, while the effect of insulin is markedly increased in the transfected cells; (ii) PAO does not affect the tyrosine phosphorylation of the insulin receptor; (iii) the tyrosine kinase activity of the insulin receptor toward exogenous substrates is not increased by PAO. Since PAO appears to act on glucose transport by a different mechanism than insulin, we have compared the effect of PAO and insulin on tyrosine phosphorylation of cellular proteins. Using Western blot analysis we did not detect common substrates in PAO- and insulin-treated cells. However, we found in cell extracts from both PAO- and insulin-treated cells a 50-kDa protein that is immunoprecipitated by antiphosphotyrosine antibody. In addition, PAO activates a cytosolic tyrosine kinase capable of poly(Glu/Tyr) phosphorylation. As a whole, our data suggest that the 50-kDa protein found in cells incubated with PAO and insulin could be the convergence point of the insulin and PAO signaling pathways.
Ballotti
R
R
INSERM U145, Faculté de Médecine, Nice, France.
Tartare
S
S
Chauvel
A
A
Scimeca
J C
JC
Alengrin
F
F
Filloux
C
C
Van Obberghen
E
E
D016475:3T3 Cells; D000818:Animals; D001152:Arsenicals / Q000494:pharmacology*; D002460:Cell Line; D002474:Cell-Free System; D003600:Cytosol / Q000201:enzymology; D003847:Deoxyglucose / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D051379:Mice; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000302:isolation & purification / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D011972:Receptor, Insulin / Q000378:metabolism
D001152:Arsenicals; D007328:Insulin; D011506:Proteins; C029341:oxophenylarsine; D003847:Deoxyglucose; D008715:Methionine; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D010749:Phosphoprotein Phosphatases; D005947:Glucose