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Bautista, S. J.

Publications and source records attributed to Bautista, S. J..

3 recordsLinked to original sources

Control of GSK3beta nuclear localization by amino acid signaling requires GATOR1 but is mTORC1 independent

The availability of certain amino acids regulates cell survival, proliferation, growth, differentiation, and other cellular functions. Sensing of amino acids that converges on the GATOR1 and GATOR2 complexes supports activation of mTORC1 during amino acid replete conditions. Whether amino acid-derived cues regulate additional pathways remains poorly understood. We uncover that amino acid sensing involving GATOR1 and GATOR2 regulates the cellular localization of glycogen synthase kinase 3{beta} (GSK3{beta}). GATOR1 is required to recruit a subset of GSK3{beta} to the lysosome selectively in the presence of amino acids. In addition, while under nutrient replete conditions GSK3{beta} is largely cytosolic, amino acid starvation drives a portion of GSK3{beta} into the nucleus. Acute replenishment of specific amino acids in starved cells triggered nuclear exit of GSK3{beta}. This amino acid-stimulated GSK{beta} nuclear exit required GATOR1 and GATOR2 but was independent of mTORC1 and its activating RagA/B GTPases. This suggests that GATOR1 has a function that diverges from control of mTORC1 to regulate the nucleocytoplasmic shuttling of GSK3{beta}. Furthermore, experimental restriction of GSK3{beta} to the cytoplasm decreased cell survival in amino acid deficient conditions. This suggests that control of GSK3{beta} nuclear localization by GATOR-dependent signals represents a cellular adaptation to metabolic stress that supports cell survival.

cell biology↗

Targeted enhancement of flotillin-dependent endocytosis augments cellular uptake and impact of cytotoxic drugs

Cellular uptake is limiting for the efficacy of many cytotoxic drugs used to treat cancer. Identifying endocytic mechanisms that can be modulated with targeted, clinically-relevant interventions is important to enhance the efficacy of various cancer drugs. We identify that flotillin-dependent endocytosis can be targeted and upregulated by ultrasound and microbubble (USMB) treatments to enhance uptake and efficacy of cancer drugs such as cisplatin. USMB involves targeted ultrasound following administration of encapsulated microbubbles, used clinically for enhanced ultrasound image contrast. USMB treatments robustly enhanced internalization of the molecular scaffold protein flotillin, as well as flotillin-dependent fluid-phase internalization, a phenomenon dependent on the protein palmitoyltransferase DHHC5 and the Src-family kinase Fyn. USMB treatment enhanced DNA damage and cell killing elicited by the cytotoxic agent cisplatin in a flotillin-dependent manner. Thus, flotillin-dependent endocytosis can be modulated by clinically-relevant USMB treatments to enhance drug uptake and efficacy, revealing an important new strategy for targeted drug delivery for cancer treatment.

cell biology↗

mTORC1 controls glycogen synthase kinase 3β nuclear localization and function

Glycogen synthase kinase 3{beta} (GSK3{beta}) phosphorylates and regulates a wide range of substrates involved in diverse cellular functions. Some GSK3{beta} substrates, such as c-myc and snail, are nuclear-resident transcription factors, suggesting possible control of GSK3{beta} function by regulation of its nuclear localization. Inhibition of mechanistic target of rapamycin (mTORC1) led to partial redistribution of GSK3{beta} from the cytosol to the nucleus, and GSK3{beta}-dependent reduction of the expression of c-myc and snail. mTORC1 is controlled by metabolic cues, such as by AMP-activated protein kinase (AMPK) or amino acid abundance. Indeed AMPK activation or amino acid deprivation promoted GSK3{beta} nuclear localization in an mTORC1-dependent manner. GSK3{beta} was detected in several distinct endomembrane compartments, including lysosomes. Consistently, disruption of late endosomes/lysosomes through perturbation of Rab7 resulted in loss of GSK3{beta} from lysosomes, and enhanced GSK3{beta} nuclear localization as well as GSK3{beta}-dependent reduction of c-myc levels. This indicates that GSK3{beta} nuclear localization and function is suppressed by mTORC1, and suggests a new link between metabolic conditions sensed by mTORC1 and GSK3{beta}-dependent regulation of transcriptional networks controlling biomass production.\n\nSummary statement (15-30 words)GSK3{beta} nuclear localization and function is negatively regulated by the metabolic and mitogenic sensor mTORC1. mTORC1 control of GSK3{beta} localization requires Rab7 and lysosomal membrane traffic.

cell biology↗