TFEB links autophagy to lysosomal biogenesis. The study shows that starvation activates a transcriptional program that controls key steps of the autophagic pathway, including autophagosome formation, autophagosome-lysosome fusion, and substrate degradation. The transcription factor TFEB, a master gene for lysosomal biogenesis, coordinates this program by driving the expression of autophagy and lysosomal genes. TFEB's nuclear localization and activity are regulated by serine phosphorylation mediated by extracellular signal-regulated kinase 2 (ERK2), whose activity is controlled by extracellular nutrient levels. This mechanism regulates autophagy by controlling the biogenesis and partnership of two distinct cellular organelles. Macro-autophagy is an evolutionary conserved mechanism that targets intracytoplasmic material to lysosomes, providing energy during nutrient starvation. Autophagy activation during starvation is negatively regulated by mTORC1, whose activity depends on cellular energy needs. The study found that starvation induces the transcription of several autophagy genes, while inhibition of mTORC1 does not, suggesting an alternative transcriptional mTORC1-independent regulation of autophagy. The study tested whether TFEB, a transcription factor that controls lysosomal biogenesis, also regulates autophagy. Overexpression of TFEB in HeLa cells significantly increased the number of autophagosomes detected by immunofluorescence and immunoblotting of the LC3 protein. Similar results were obtained in other cell types. This increase persisted in cells treated with lysosomal inhibitors, indicating that TFEB activates autophagosome formation. RNA interference of TFEB in HeLa cells resulted in decreased levels of LC3-II, indicating that TFEB is involved in the biogenesis of autophagosomes and lysosomes. The study also measured the rate of delivery of autophagosomes to lysosomes using an RFP-GFP tandem tagged LC3 protein, which discriminates early autophagic organelles from acidified autophagolysosomes. The number of autophagolysosomes was higher in TFEB overexpressing cells, indicating that TFEB enhances the autophagic flux. The study analyzed the mRNA levels of 51 genes involved in several steps of the autophagic pathway. The enhancement of the expression levels of autophagy genes in cells overexpressing TFEB was very similar to that of starved cells. Eleven of the analyzed genes were significantly up-regulated after TFEB overexpression, while they were down-regulated after TFEB silencing. These genes are known to play a role in different steps of autophagy and appear to be direct targets of TFEB. The study also showed that TFEB is involved in the transcriptional regulation of starvation-induced autophagy. The findings suggest that TFEB is involved in the transcriptional regulation of starvation-induced autophagy. The study also identified a novel transcriptional mechanism that controlsTFEB links autophagy to lysosomal biogenesis. The study shows that starvation activates a transcriptional program that controls key steps of the autophagic pathway, including autophagosome formation, autophagosome-lysosome fusion, and substrate degradation. The transcription factor TFEB, a master gene for lysosomal biogenesis, coordinates this program by driving the expression of autophagy and lysosomal genes. TFEB's nuclear localization and activity are regulated by serine phosphorylation mediated by extracellular signal-regulated kinase 2 (ERK2), whose activity is controlled by extracellular nutrient levels. This mechanism regulates autophagy by controlling the biogenesis and partnership of two distinct cellular organelles. Macro-autophagy is an evolutionary conserved mechanism that targets intracytoplasmic material to lysosomes, providing energy during nutrient starvation. Autophagy activation during starvation is negatively regulated by mTORC1, whose activity depends on cellular energy needs. The study found that starvation induces the transcription of several autophagy genes, while inhibition of mTORC1 does not, suggesting an alternative transcriptional mTORC1-independent regulation of autophagy. The study tested whether TFEB, a transcription factor that controls lysosomal biogenesis, also regulates autophagy. Overexpression of TFEB in HeLa cells significantly increased the number of autophagosomes detected by immunofluorescence and immunoblotting of the LC3 protein. Similar results were obtained in other cell types. This increase persisted in cells treated with lysosomal inhibitors, indicating that TFEB activates autophagosome formation. RNA interference of TFEB in HeLa cells resulted in decreased levels of LC3-II, indicating that TFEB is involved in the biogenesis of autophagosomes and lysosomes. The study also measured the rate of delivery of autophagosomes to lysosomes using an RFP-GFP tandem tagged LC3 protein, which discriminates early autophagic organelles from acidified autophagolysosomes. The number of autophagolysosomes was higher in TFEB overexpressing cells, indicating that TFEB enhances the autophagic flux. The study analyzed the mRNA levels of 51 genes involved in several steps of the autophagic pathway. The enhancement of the expression levels of autophagy genes in cells overexpressing TFEB was very similar to that of starved cells. Eleven of the analyzed genes were significantly up-regulated after TFEB overexpression, while they were down-regulated after TFEB silencing. These genes are known to play a role in different steps of autophagy and appear to be direct targets of TFEB. The study also showed that TFEB is involved in the transcriptional regulation of starvation-induced autophagy. The findings suggest that TFEB is involved in the transcriptional regulation of starvation-induced autophagy. The study also identified a novel transcriptional mechanism that controls