Evaluating the mTOR Pathway in Physiological and Pharmacological Settings.
Mammalian/mechanistic target of rapamycin (mTOR) is an evolutionarily conserved genuine protein kinase, which phosphorylates serine/threonine in response to growth factors and nutrients. It functions as a catalytic core in two distinct multiprotein complexes: mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). mTORC1 promotes cell growth and proliferation by positively regulating translation, transcription, and lipid biosynthesis in response to growth factors and amino acids, whereas it inhibits autophagy, an essential degradation and recycling pathway. mTORC2 regulates cell survival and cytoskeleton organization. Mechanistic insights into the function and regulation of mTOR complexes have been provided in various experimental settings and monitoring mTOR activity has been a most valuable way to judge whether levels of environmental cues such nutrients and growth factors can satisfy cellular needs for cell growth, proliferation, and autophagic response. Here, we describe useful methods to access mTOR activity in different experimental settings.