In vivo characterisation of the human UCP3 gene minimal promoter in mice tibialis anterior muscles.

ABSTRACT

Transcriptional mechanisms controlling human UCP3 gene expression in skeletal muscle remain poorly understood. Experiments based on plasmid electrotransfer into tibialis anterior muscle of C57/BL6 male mice were set up in order to functionally analyze the hUCP3 gene promoter. These transfection experiments showed that a 6300 bp region upstream of the transcription initiation site was sufficient to mediate maximal promoter activity. Further analyses with a series of 5(')-deleted constructs demonstrated that the hUCP3 gene minimal promoter was located between nucleotides -284 and -40. Furthermore, an essential region was identified between nucleotides -284 and -224. The analysis of this region revealed a putative response element for PPAR located between nucleotides -281 and -269. Finally, mutations of potential cis-acting elements within the hUCP3 minimal promoter showed the presence of two TATA boxes (-198/-194 and -45/-41) required for constitutive UCP3 gene expression. To our knowledge, this is the first time that molecular characterization of the UCP3 promoter has been achieved using an in vivo experimental model.