The Pxmp2 and PoleI genes are linked by a bidirectional promoter in an evolutionary conserved fashion

• Published in: Gene 2003-08, Vol.313, p.119-126
• Main Authors: Otte, David M, Schwaab, Ulrike, Lüers, Georg H
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Bidirectional promoter; Blotting, Northern; Cell Line; CHO Cells; Chromosomes, Mammalian - genetics; Conserved Sequence - genetics; Cricetinae; DNA Polymerase II - genetics; Evolution, Molecular; Gene Expression; Genetic Linkage; Luciferases - genetics; Luciferases - metabolism; Membrane Proteins - genetics; Mice; Molecular Sequence Data; PoleI; Poly-ADP-Ribose Binding Proteins; Promoter Regions, Genetic - genetics; Pxmp2; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sequence Homology, Nucleic Acid; Transfection
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(03)00667-X

Pxmp2 is the most abundant peroxisomal membrane protein in higher eukaryotes. Its expression is tissue-specific with highest levels of expression in liver, kidney and heart tissue. We have analysed the 5′-flanking genomic region of the murine Pxmp2 gene and we found, that the first exon of the gene encoding the DNA polymerase ε ( PoleI) was localized adjacent to the first exon of the Pxmp2 gene in head to head orientation. Both genes were separated by only 393 bp containing a CpG island with numerous binding sites for Sp1. A TATA box, however, was lacking. Northern blot analysis revealed that both genes were expressed differently, indicating that their expression was regulated independently. We have analysed the promoter activity of the small genomic fragment separating the Pxmp2 and PoleI genes using luciferase as a reporter molecule in transient transfection assays. The small genomic fragment was a functional promoter, controlling gene expression regardless of its orientation. Promoter activity was 60-70% compared with the activity of the strong CMV promoter. The Pxmp2 and PoleI genes were also linked on the human and rat genome. Furthermore, the sequence of the intergenic fragment was highly conserved among these species. Thus, the small intergenic fragment is probably the common basic element of two independently regulated promoters.