TY  - JOUR
AU  - Marin Zapata, Paula Andrea
AU  - Beese, Carsten Jörn
AU  - Jünger, Anja
AU  - Dalmasso, Giovanni
AU  - Brady, Nathan R
AU  - Hamacher-Brady, Anne
TI  - Time course decomposition of cell heterogeneity in TFEB signaling states reveals homeostatic mechanisms restricting the magnitude and duration of TFEB responses to mTOR activity modulation.
JO  - BMC cancer
VL  - 16
IS  - 1
SN  - 1471-2407
CY  - London
PB  - BioMed Central
M1  - DKFZ-2017-06076
SP  - 355
PY  - 2016
AB  - TFEB (transcription factor EB) regulates metabolic homeostasis through its activation of lysosomal biogenesis following its nuclear translocation. TFEB activity is inhibited by mTOR phosphorylation, which signals its cytoplasmic retention. To date, the temporal relationship between alterations to mTOR activity states and changes in TFEB subcellular localization and concentration has not been sufficiently addressed.mTOR was activated by renewed addition of fully-supplemented medium, or inhibited by Torin1 or nutrient deprivation. Single-cell TFEB protein levels and subcellular localization in HeLa and MCF7 cells were measured over a time course of 15 hours by multispectral imaging cytometry. To extract single-cell level information on heterogeneous TFEB activity phenotypes, we developed a framework for identification of TFEB activity subpopulations. Through unsupervised clustering, cells were classified according to their TFEB nuclear concentration, which corresponded with downstream lysosomal responses.Bulk population results revealed that mTOR negatively regulates TFEB protein levels, concomitantly to the regulation of TFEB localization. Subpopulation analysis revealed maximal sensitivity of HeLa cells to mTOR activity stimulation, leading to inactivation of 100 
KW  - Basic Helix-Loop-Helix Leucine Zipper Transcription Factors (NLM Chemicals)
KW  - TFEB protein, human (NLM Chemicals)
KW  - MTOR protein, human (NLM Chemicals)
KW  - TOR Serine-Threonine Kinases (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:27268034
C2  - pmc:PMC4896000
DO  - DOI:10.1186/s12885-016-2388-9
UR  - https://inrepo02.dkfz.de/record/131000
ER  -