INSULIN-RECEPTOR SUBSTRATE-1 IS PHOSPHORYLATED BY THE SERINE KINASE-ACTIVITY OF PHOSPHATIDYLINOSITOL 3-KINASE

Insulin receptor substrate (IRS) 1, which is tyrosine phosphorylated i n response to insulin, presents multiple serine/threonine phosphorylat ion sites. To search for a serine kinase activity towards IRS 1, immun oprecipitates from basal or stimulated 3T3-L1 adipocytes were used in an in vitro kinase assay. When IRS 1 was isolated from insulin-treated cells, serine phosphorylation of IRS 1 occurred, which we attribute t o the kinase activity of the phosphatidylinositol 3-kinase (PI3-kinase ). Importantly, in an in vitro reconstitution assay, an excess of the PI3-kinase subunit prevents this phosphorylation. Together, our result s suggest that following insulin stimulation, PI3-kinase associates wi th IRS 1, allowing for its serine phosphorylation. This phosphorylatio n event could play a role in the modulation of insulin signalling.