CCAAT/enhancer-binding protein beta (C/EBP beta) is expressed early during the adipocyte differentiation program and plays an important role in this process. In an attempt to identify novel proteins that interact with C/EBP beta, we performed a yeast two-hybrid screen with a preadipocyte cDNA library and identified a new co-regulator, delta-interacting protein A (DIPA). DIPA mRNA is expressed during adipocyte differentiation of clonal cell lines. DIPA interacts with C/EBP beta and -delta proteins in intact cells and inhibits their transcriptional activity but not that of C/EBP alpha. Stable overexpression of DIPA in preadipocytes partially inhibits adipocyte differentiation, whereas its gene silencing enhances this process. DIPA and C/EBP beta co-localize in the nucleus, and overexpression of DIPA in preadipocytes results in a partial inhibition of the mitotic clonal expansion which is critical for differentiation. Thus, DIPA is a novel partner of C/EBP beta that down-regulates early events of adipogenesis.