Molecular characterization of atrogin-1/F-box protein-32 (FBXO32) and F-box protein-25 (FBXO25) in rainbow trout (Oncorhynchus mykiss): Expression across tissues in response to feed deprivation.

The characteristic increase in protein catabolism during muscle atrophy is largely the result of an increase in E3 ubiquitin ligase expression, specifically that of atrogin-1, or FBXO32, which functions to polyubiquitinate proteins. In rainbow trout, the cDNA sequences of two E3 ubiquitin ligase F-box proteins, FBXO32 and FBXO25, were characterized and their expression across tissues in response to feed deprivation was determined. The cDNA sequence for FBXO32 encodes a protein 355 amino acids long and is 97% identical to the homologous protein in salmon, 85% to zebrafish and 72% identical to both human and mouse. The cDNA for FBXO25 encodes a protein 356 amino acids in length that is 98% identical to the homologous protein in salmon, 84% to zebrafish, and 75% to human. After 28days of feed deprivation, FBXO32 expression increased by approximately 13-fold, 3-fold, and 5-fold in white muscle, red muscle, and intestine, respectively (P<0.05). Expression of FBXO32 and FBXO25 in kidney decreased 0.3-fold and 0.2-fold, respectively, and FBXO25 expression decreased by 0.2-fold in liver (P<0.05). These results indicate that these protein sequences are conserved and suggest that the up-regulation of FBXO32 is associated with skeletal and smooth muscle atrophy that occurs during fasting.