Preparing your results

Our searching services are busy right now. Your search will reload in five seconds.

X
Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.

Immunopurified mammalian target of rapamycin phosphorylates and activates p70 S6 kinase alpha in vitro.

http://www.ncbi.nlm.nih.gov/pubmed/10567431

p70 S6 kinase alpha (p70alpha) is activated in vivo through a multisite phosphorylation in response to mitogens if a sufficient supply of amino acids is available or to high concentrations of amino acids per se. The immunosuppressant drug rapamycin inhibits p70alpha activation in a manner that can be overcome by coexpression of p70alpha with a rapamycin-resistant mutant of the mammalian target of rapamycin (mTOR) but only if the mTOR kinase domain is intact. We report here that a mammalian recombinant p70alpha polypeptide, extracted in an inactive form from rapamycin-treated cells, can be directly phosphorylated by the mTOR kinase in vitro predominantly at the rapamycin-sensitive site Thr-412. mTOR-catalyzed p70alpha phosphorylation in vitro is accompanied by a substantial restoration in p70alpha kinase activity toward its physiologic substrate, the 40 S ribosomal protein S6. Moreover, sequential phosphorylation of p70alpha by mTOR and 3-phosphoinositide-dependent protein kinase 1 in vitro resulted in a synergistic stimulation of p70alpha activity to levels similar to that attained by serum stimulation in vivo. These results indicate that mTOR is likely to function as a direct activator of p70 in vivo, although the relative contribution of mTOR-catalyzed p70 phosphorylation in each of the many circumstances that engender p70 activation remains to be defined.

Pubmed ID: 10567431 RIS Download

Mesh terms: 3-Phosphoinositide-Dependent Protein Kinases | Cell Line | Culture Media, Serum-Free | Enzyme Activation | Humans | Immunoblotting | Mitogens | Mutation | Phosphorylation | Phosphotransferases (Alcohol Group Acceptor) | Precipitin Tests | Protein Kinases | Protein-Serine-Threonine Kinases | Recombinant Fusion Proteins | Ribosomal Protein S6 Kinases | TOR Serine-Threonine Kinases