Carbohydrate regulation of hepatic gene expression. Evidence against a role for the upstream stimulatory factor

Hepatic expression of the genes encoding L-type pyruvate kinase (L-PK) and S14 is induced in rats upon feeding them a high carbohydrate, low fat diet. A carbohydrate response element (ChoRE) containing two CACGTG-type E boxes has been mapped in the 5'-flanking region of both of these genes. The nature of the ChoRE suggests that a member of the basic/helix-loop-helix/leucine zipper family of proteins may be responsible for mediating the response to carbohydrate. Indeed, the upstream stimulatory factor (USF), a ubiquitous basic/helix-loop-helix/leucine zipper protein, is present in hepatic nuclear extracts and binds to the ChoREs of L-PK and S14 in vitro. We have conducted experiments to determine whether USF is involved in the carbohydrate-mediated regulation of L-PK and S14. For this purpose, dominant negative forms of USF that are capable of heterodimerizing with endogenous USF but not of binding to DNA were expressed in primary hepatocytes. Expression of these forms did not block either S14 or L-PK induction by glucose. In addition, we have constructed mutant ChoREs that retain their carbohydrate responsiveness but have lost the ability to bind USF. Together, these data suggest that USF is not the carbohydrate-responsive factor that stimulates S14 and L-PK expression and that a distinct hepatic factor is likely to be responsible for the transcriptional response.