Drosophila melanogaster activating transcription factor 4 regulates glycolysis during endoplasmic reticulum stress.
Bottom Line: The unfolded protein response transcription factor Atf4 was necessary for the up-regulation of glycolytic enzymes and Lactate dehydrogenase (Ldh).Finally, flies up-regulated Ldh and produced more lactate when subjected to ER stress.Together, these results suggest that Atf4 mediates a shift from a metabolism based on oxidative phosphorylation to one more heavily reliant on glycolysis, reminiscent of aerobic glycolysis or the Warburg effect observed in cancer and other proliferative cells.
Affiliation: Department of Biology, University of Utah, Salt Lake City, Utah 84112.Show MeSH
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Mentions: We transfected S2 cells with 2 μg of plasmid using Cellfectin II (Invitrogen). For polyclonal stable cell lines (as in Figure 3D and Figure 5A), we cotransfected our expression plasmids (1.8 μg) with a hygromycin or puromycin resistance plasmid (0.2 μg) and selected for resistant cells. For Atf4 overexpression studies (Figure 3D), we induced expression with CuSO4 (250 μM, 36 hr) before collecting RNA samples. For promoter constructs, we treated control cells transfected with pMT-GFP with CuSO4 (250 μM, 16 hr) before inducing ER stress with DTT as described above. Finally, for transiently transfected cells (Figure 5B), we allowed cells to recover overnight before inducing ER stress.
Affiliation: Department of Biology, University of Utah, Salt Lake City, Utah 84112.