Article

Diabetologia

, Volume 46, Issue 11, pp 1532-1542

First online:

MAP kinases and mTOR mediate insulin-induced phosphorylation of Insulin Receptor Substrate-1 on serine residues 307, 612 and 632

  • P. GualAffiliated withINSERM U568 and IFR50, Faculté de Médecine
  • , T. GrémeauxAffiliated withINSERM U568 and IFR50, Faculté de Médecine
  • , T. GonzalezAffiliated withINSERM U568 and IFR50, Faculté de Médecine
  • , Y. Le Marchand-BrustelAffiliated withINSERM U568 and IFR50, Faculté de Médecine Email author 
  • , J.-F. TantiAffiliated withINSERM U568 and IFR50, Faculté de Médecine

Abstract

Aim/hypothesis

Insulin-induced IRS-1 serine phosphorylation could be physiologically important to regulate insulin action. In a hyperinsulinaemic state such as obesity or Type 2 diabetes, this phosphorylation could be modified and exacerbate insulin resistance. We aimed at identifying serine residues in IRS-1 phosphorylated in response to insulin stimulation and at determining the involved kinases.

Methods

3T3-L1 adipocytes, muscle and adipose tissue of mice were subjected to Western Blot analysis with phosphospecific antibodies to identify phosphorylation sites in IRS-1 following insulin treatment. Pharmacological inhibitors were used to determine the serine kinases involved in this phosphorylation.

Results

In 3T3-L1 adipocytes, insulin promoted the phosphorylation of serine 307, 612 and 632 with Serine612/632 more rapidly phosphorylated than Serine307. Insulin-induced phosphorylation of Serine307 was dependent on the activation of a PI 3-kinase/mTOR pathway. The phosphorylation of Serine612/632 required the activation of the MAP kinase pathway following short-term insulin stimulation and activation of the PI 3-kinase/mTOR pathway following prolonged insulin stimulation. Phosphorylation of Serine307 and Serine632 occurred in vivo in skeletal muscle and white adipose tissue of mice injected with insulin and was dependent on the activation of mTOR. Moreover, inhibition of mTOR led to a persistent PI 3-kinase activation by insulin.

Conclusion/Interpretation

Insulin-induced IRS-1 serine phosphorylation is a complex process involving different sites and kinases. This complexity could be physiologically important to accurately regulate insulin signalling. Abnormal phosphorylation of these serine residues in hyperinsulinaemic state could participate in the down-regulation of insulin signalling.

Keywords

IRS-1 PI 3-kinase insulin resistance diabetes serine phosphorylation muscles adipose tissue 3T3-L1 adipocytes rapamycin