biostudies-literatureUnknown65(11)Cansby EMetabolic dysfunction-associated steatotic liver disease has emerged as a leading global cause of chronic liver disease. Our recent translational investigations have shown that the STE20-type kinases comprising the GCKIII subfamily-MST3, STK25, and MST4-associate with hepatic lipid droplets and regulate ectopic fat storage in the liver; however, the mode of action of these proteins remains to be resolved. By comparing different combinations of the silencing of MST3, STK25, and/or MST4 in immortalized human hepatocytes, we found that their single knockdown results in a similar reduction in hepatocellular lipid content and metabolic stress, without any additive or synergistic effects observed when all three kinases are simultaneously depleted. A genome-wide yeast two-hybrid screen of the humJournal of lipid research100669https://www.ebi.ac.uk/biostudies/studies/S-EPMC11602991biostudies-literatureGCKIII kinases control hepatocellular lipid homeostasis via shared mode of action.PMC11602991Xia YMahlapuu MHenricsson MSvensson LTAsiedu BAndersson EHoy AJSaghaleyni RBluher MCaputo MCansby EfalseGCKIII kinases control hepatocellular lipid homeostasis via shared mode of action.Metabolic dysfunction-associated steatotic liver disease has emerged as a leading global cause of chronic liver disease. Our recent translational investigations have shown that the STE20-type kinases comprising the GCKIII subfamily-MST3, STK25, and MST4-associate with hepatic lipid droplets and regulate ectopic fat storage in the liver; however, the mode of action of these proteins remains to be resolved. By comparing different combinations of the silencing of MST3, STK25, and/or MST4 in immortalized human hepatocytes, we found that their single knockdown results in a similar reduction in hepatocellular lipid content and metabolic stress, without any additive or synergistic effects observed when all three kinases are simultaneously depleted. A genome-wide yeast two-hybrid screen of the hum2024-01-01T00:00:00Z2024 Nov2026-06-02T23:10:13.226Z2025-04-04T00:46:24.643ZS-EPMC116029913939579110.1016/j.jlr.2024.100669