Supramolecular Assembly of GSK3α as a Cellular Response to Amino Acid Starvation
Description
This data submission contains the raw confocal immunofluorescence microscopy and Western blot images for our manuscript, which describes the following: The tolerance of amino acid starvation is fundamental to robust cellular fitness. Asparagine depletion is lethal to some cancer cells, a vulnerability exploited clinically using asparaginase. We report that resistance to asparagine starvation is uniquely dependent on an N-terminal low-complexity domain of GSK3α lacking in its paralog GSK3β. In response to depletion of specific amino acids, including asparagine, leucine or valine, this domain mediates supramolecular assembly of GSK3α with ubiquitin-proteasome system components in spatially sequestered cytoplasmic bodies. This effect is independent of mTORC1 or GCN2. In normal cells, GSK3α promotes survival during essential amino acid starvation. In human leukemia, GSK3α body formation predicts asparaginase resistance, and sensitivity to asparaginase combined with a GSK3α inhibitor. We propose that GSK3α body formation provides a cellular mechanism to maximize the catalytic efficiency of proteasomal protein degradation in response to amino acid starvation, an adaptive response co-opted by cancer cells for asparaginase resistance.
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Steps to reproduce
Full methods are included in our manuscript. Note that each microscopy image file contains multiple images from the different fluorescent channels utilized, the different fluorophores can be seen using ImageJ.