Interaction of insulin receptor substrate-2 (IRS-2) with the insulin and insulin-like growth factor I receptors - Evidence for two distinct phosphotyrosine-dependent interaction domains within IRS-2

Insulin receptor substrate 2 (IRS-2) has recently been shown to be a substrate of the insulin receptor (IR), In this study we utilize the yeast two-hybrid system and assays of in vitro interaction to demonstrate that IRS-2 interacts directly with the IR and the insulin-like growth factor I receptor, We show that, like IRS-1, the region of IRS-2 that contains the putative phosphotyrosine binding and SAIN elements (188-591) is sufficient for receptor interaction and that this interaction is dependent upon the NPX(p)Y (where (p)Y is phosphotyrosine) motifs within the juxtamembrane domains of the receptors, In addition to this amino-terminal NPX(p)Y-binding domain, an additional domain of strong interaction was identified in the central region of IRS-2 and was localized between amino acids 591 and 733, This interaction was found to be dependent upon receptor phosphorylation but was NPX(p)Y-independent, This region does not appear to have either an SH2 or a phosphotyrosine binding domain, Both of the interactions could also be demonstrated in vitro using IRS-2 glutathione S-transferase fusion proteins, We conclude that IRS-2, unlike IRS-1, can interact with tyrosine phosphorylated receptors such as the IR and insulin-like growth factor I receptor via multiple independent binding motifs, Our findings suggest the existence of a previously unidentified phosphotyrosine dependent binding domain within the central region of IRS-2.