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

• Published in: Biochemical and biophysical research communications 2003-11, Vol.311 (3), p.583-591
• Main Authors: Riquet, Franck B, Rodriguez, Marianne, Guigal, Nolwen, Dromaint, Sandra, Naime, Isabelle, Boutin, Jean A, Galizzi, Jean-Pierre
• Format: Article
• Language: English
• Subjects: Animals; Carrier Proteins - chemistry; Carrier Proteins - genetics; CHO Cells; COS Cells; Cricetinae; DNA - metabolism; Electroporation; Gene Deletion; Genes, Reporter; Human UCP3; Humans; Ion Channels; Male; Mice; Mice, Inbred C57BL; Mitochondrial Proteins; Models, Genetic; Mouse; Muscle; Muscle, Skeletal - metabolism; Muscles - metabolism; Mutagenesis, Site-Directed; Mutation; Plasmids - metabolism; Promoter; Promoter Regions, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Time Factors; Transcription, Genetic; Transfection; UCP3 gene; Uncoupling Protein 3
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2003.10.034

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.