Transient induction of ENC-1, a Kelch-related actin-binding protein, is required for adipocyte differentiation

In an attempt to study molecules that play a regulatory role early in adipocyte differentiation, we identified by differential display ENC-1, a Drosophila kelch-related protein. ENC-1 colocalizes with actin filaments. ENC-1 is expressed in adipose tissue, specifically in the adipose-derived stroma-vascular fraction. ENC-1 mRNA levels are transiently increased 8-12-fold early in in vitro adipocyte differentiation of primary cells of the adipose-derived stroma-vascular fraction and of 3T3-L1 cells. Treatment with the adipogenic inducers dexamethasone and methylisobutylxanthine causes an increase in ENC-1 mRNA levels specifically in preadipocytes, and methylisobutylxanthine is the main effector of ENC-1 expression. The induction of ENC-1 precedes expression of the transcription factors, peroxisome proliferator-activated receptor (PPARgamma) and CCAAT/enhancer-binding protein (C/EBPalpha), and other adipocyte markers including adipocyte fatty acid-binding protein. The ENC-1 induction correlates with the subsequent differentiation of primary stroma-vascular cells into adipocytes. Furthermore, decreasing the endogenous ENC-1 levels by stable antisense transfection, thereby preventing the transient induction, effectively inhibits 3T3-L1 adipocyte differentiation. Overall, these studies indicate that ENC-1, an actin-binding protein, plays a regulatory role early in adipocyte differentiation when cytoskeletal reorganization and cell shape change from fibroblastic preadipocytes to spherical adipocytes occur.