Transcription of genes encoding enzymes required for lipogenesis is induced in hepatocytes in response to elevated glucose metabolism. We have previously mapped the carbohydrate-response elements (ChoREs) of the rat liver-type pyruvate kinase (L-PK) and S14 genes and found them to share significant sequence similarity. However, progress in unraveling this signaling pathway has been hampered due to the difficulty in identifying the key factor(s) that bind to these ChoREs. To gain further insight into the nature of the carbohydrate-responsive transcription factor, the glucose regulatory sequences from the mouse S14 gene were examined in primary hepatocytes. Three elements were found to be essential for supporting the glucose response: a thyroid hormone-response element between -1522 and -1494, an accessory factor site between -1421 and -1392, and the ChoRE between -1450 and -1425. Of these, only the accessory factor site was conserved between the rat and mouse S14 genes. Investigation of the ChoRE sequence indicated that two half E box motifs are critical for the response to glucose. Electrophoretic mobility shift assays revealed a complex formed between the mouse S14 ChoRE and liver nuclear proteins. This complex was also formed by ChoREs from the rat S14 and L-PK genes but not by mutants of these sites that are inactive in supporting the glucose response. These results suggest the presence of a novel transcription factor complex that mediates the glucose-regulated transcription of S14 and L-PK genes.
|Number of pages||8|
|Journal||Journal of Biological Chemistry|
|Publication status||Published - 2000 Feb 18|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology