Exploration of a liver-specific, glucose/insulin-responsive promoter in transgenic mice.

The functional role of the different sites binding transcriptional factors on the tissue-specific, glucose-responsive promoter of the L type pyruvate kinase gene (L-PK) has been investigated in transgenic mice. These sites are able to bind, from 3' to 5', HNF1, NF1, HNF4, and MLTF/USF, respectively. We have compared the level of chloramphenicol acetyltransferase reporter transgene expression when driven by a L-PK promoter fragment of either -96 base pairs (bp) (containing only the HNF1 binding site) or -150 bp (lacking the MLTF/USF binding site) or driven by a -183-bp L-PK promoter fragment with or without the NF1 binding site. Our results demonstrate that: 1) HNF1 alone is not sufficient to promote an efficient L-PK gene transcription in vivo; 2) with only binding sites for HNF1, NF1, and HNF4, though the tissue-specific pattern of expression is respected, the level of the gene transcription is low and the hormonal control is lost; 3) the MLTF/USF binding site is the target of the hormonal control, required for both positive response to carbohydrates and negative response to glucagon; 4) the role of NF1 in the promoter activity could be to negatively modulate the L-PK gene expression in the different tissues, without interfering with the glucose and hormone responsiveness.