Determine the role of P38 kinase in insulin signaling

IRS1 (Insulin Receptor Substrate 1) is the main adaptor of Insulin Receptor (IR) and is responsible for pozitive and negative regulation of insuling signaling. During insulin signaling ligand-bound IR is phosphorylated at tyrosine residues via auto-phosphorylation, then, IRS1 binds to these phosphotyrosine through its SH2 domain. IR-bound IRS1 is phosphorylated by IR at YxxM motifs, four of which were localized at the C-terminal. These phosphorylation sites are binding sites of p85 α regulatory subunit of PI3K YXXM-bound PI3Kp85α also binds to PI3Kp110α and activates it. Activated PI3Kp110α generates PIP3 from PIP2, and therefore, activates PDK1. Activated PDK1 activates AKT by phosphorylating it at S308. Activated AKT regulates glucose-uptake by activating Glucose transporters GLUT1 and GLUT4. Although tyrosine phosphorylation of IRS1 positively stimulates insuling signaling its phosphorylations at serine/treonin (S/T) residues negatively regulates insulin signaling. For this reason, identification of enzymes responsible for S/T phosphorylations on IRS1 is important for elucidation of mechanisms for insulin signaling. Phosphorylations of IRS1 on S/T residues seem to be increased under stres conditions such as LPS, osmotic stress, ER-stress, UV and ionizing radiation and this is one of the main reason for "Insulin Resistance/Type 2 diabetes". These inducers also activate p38 MAPK. In this respect, when we analysed the amino acid sequence of human IRS1, we found GYMPMS616P, DYMPMS636P, GYMMMS666P, DYMNMS736P motifs which are phosphorylation sides of p38 MAPK. In our study we showed that these indicated sites were phosphorylated by p38 MAPK is activated by anisomycin. To further analyse the significance of phosphorylation of these sites, we converted above residues to unphosphorylated aminoacid Alanine or phosphorylation mimicking negatively charged Glutamic acid. Our results suggested that p38 MAPK has an important role of development of insülin resistance and cell proliferation through IRS1 Ser/Thr phosphorylations. The effects of p38 MAPK on glucose uptake and cell proliferation can be positive or negative depends on its phosphorylation sides of IRS1. Under the light of these findings, we suggested that IRS1 Ser/Thr phosphorylations by p38 MAPK showed specific effect on insulin signaling depends on its phosphorylation sites.