Grb-2-Associated Binder-1 Is Involved in Insulin-Induced egr-1 Gene Expression through its Phosphatidylinositol 3′-Kinase Binding Site

Abstract

The Grb2-associated binder-1 (Gab1) is one of the major adapter molecules downstream of growth factor receptor signaling. Even though insulin causes tyrosine phosphorylation of Gab1, its role in insulin signaling has not been identified yet. We have demonstrated that insulin increased expression of early growth response gene-1 (egr-1), which is one of the most important transcription factors involved in cell proliferation and differentiation. In the present study, the possible role of Gab1 in insulin-induced egr-1 expression was studied using Rat1 fibroblasts expressing human insulin receptors and wildtype Gab1 (HIRc/Gab1(WT)), Gab1 with three tyrosines in the phosphatidylinositol (PI) 3′-kinase binding domain mutated to phenylalanine (HIRc/Gab1(ΔPI3K)), or histidinol resistance only(HIRc/HIS). Insulin-induced egr-1 expression in HIRc/Gab1(ΔPI3K) cells was much lower than in the other cells, as determined by Northern blot analysis. These results suggest that Gab1 is involved in the signaling pathway for insulin-induced egr-1 expression through increasing PI3′-kinase activity. The MAP kinase activity increased less with insulin treatment in HIRc/Gab1(ΔPI3K) cells than in other cells. Inhibition of MAP kinase by the MEK inhibitor completely abolished insulin-induced egr-1 expression. These results suggest that Gab1 increases MAP kinase activity through its PI3′-kinase binding site, which then leads to egr-1 expression. Our results indicate that Gab1 is involved in the control of egr-1 expression regulated by insulin.