Signaling through
mammalian target of rapamycin complex 1 (
mTORC1) is stimulated by
amino acids and
insulin.
Insulin inactivates TSC1/2, the
GTPase-activator complex for Rheb, and Rheb.
GTP activates
mTORC1. It is not clear how
amino acids regulate
mTORC1. FKBP38 (
immunophilin FK506-binding protein, 38 kDa), was recently reported to exert a negative effect on
mTORC1 function that is relieved by its binding to Rheb.
GTP. We confirm that Rheb binds wild type FKBP38, but inactive Rheb mutants showed contrasting abilities to bind FKBP38. We were unable to observe any regulation of FKBP38/mTOR binding by
amino acids or
insulin. Furthermore, FKBP38 did not inhibit
mTORC1 signaling. The
translationally controlled tumor protein (TCTP) in Drosophila was recently reported to act as the
guanine nucleotide-exchange factor for Rheb. We have studied the role of TCTP in mammalian
TORC1 signaling and its control by
amino acids. Reducing TCTP levels did not reproducibly affect
mTORC1 signaling in
amino acid-replete/
insulin-stimulated cells. Moreover, overexpressing TCTP did not rescue
mTORC1 signaling in
amino acid-starved cells. In addition, we were unable to see any stable interaction between TCTP and Rheb or
mTORC1. Accumulation of uncharged
tRNA has been previously proposed to be involved in the inhibition of
mTORC1 signaling during
amino acid starvation. To test this hypothesis, we used a Chinese hamster ovary cell line containing a temperature-sensitive mutation in leucyl-
tRNA synthetase.
Leucine deprivation markedly inhibited
mTORC1 signaling in these cells, but shifting the cells to the nonpermissive temperature for the
synthetase did not. These data indicate that uncharged
tRNA(Leu) does not switch off
mTORC1 signaling and suggest that
mTORC1 is controlled by a distinct pathway that senses the availability of
amino acids. Our data also indicate that, in the mammalian cell lines tested here, neither TCTP nor FKBP38 regulates
mTORC1 signaling.