Critical opposing roles for TFEB andTFE3 in melanoma progression

Unlike MITF-M, the melanocyte-specific isoform of MITF, TFEB, TFE3 and non-melanocyte MITF isoforms are regulated primarily by mTORC1-mediated phosphorylation at the lysosome that promotes their cytoplasmic retention. As low levels of glucose or amino acids trigger down-regulation of MITF, it is likely that on nutrient limitation some MITF functions are assumed by TFE3 and TFEB, enabling MITFLow melanoma cells to survive and proliferate. The role TFEB and TFE3 in melanoma biology and their impact on MITF-driven proliferation is poorly understood. Overall design: TFEB and TFE3 KO of 501mel melanoma cell lines were generated using guides targetting TFE3 (GGTACTGTTTCACCTGCTGC) and TFEB (AGTACCTGTCCGAGACCTAT). KO cells were clonally isolated and validated prior to experiment.