Transcription Factor EB (TFEB) is a critical regulator of lysosomal biogenesis, autophagy and energy homeostasis through regulating expression of genes belonging to the Coordinated Lysosomal Expression and Regulation network. Recently, AMP-activated protein kinase (AMPK) was reported to phosphorylate TFEB at three conserved C-terminal Ser residues (S466, S467, S469) and these phosphorylation events were essential for transcriptional activation of TFEB. In sharp contrast to this proposition, here we demonstrate that AMPK activation leads to dephosphorylation of the C-terminal sites, and that AMPK is dispensable for mTORC1-mediated/torin1-sensitive TFEB activation. We show that a synthetic peptide encompassing C-terminal Ser residues is a poor substrate of AMPK in cell-free assay. Treatment with of cells with AMPK activator (MK-8722) or mTOR inhibitor (torin1) robustly dephosphorylated TFEB not only at mTORC1-targeted N-terminal Ser sites, but also the C-terminal sites. Loss of function of AMPK abrogated MK-8722- but not torin1-induced dephosphorylation and induction of vast majority of TFEB target genes.