Human Apolipoprotein B Gene Intestinal Control Region

• Published in: Biochemistry (Easton) 2001-06, Vol.40 (23), p.6720-6730
• Main Authors: Antes, Travis J, Goodart, Sheryl A, Chen, Wei, Levy-Wilson, Beatriz
• Format: Article
• Language: English
• Subjects: Animals; apoB gene; apolipoprotein B; Apolipoproteins B - biosynthesis; Apolipoproteins B - genetics; Artificial Gene Fusion - methods; Base Composition; Base Sequence; beta-Galactosidase - biosynthesis; beta-Galactosidase - genetics; Caco-2 Cells; Deoxyribonuclease I - genetics; Enhancer Elements, Genetic; Gene Expression Regulation; Humans; Intestine, Small - metabolism; Liver - metabolism; Mice; Mice, Inbred Strains; Mice, Transgenic; Microinjections; Molecular Sequence Data; Plasmids - administration & dosage; Transgenes
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi010073a

Recently, we reported that a 315 bp enhancer, located over 55 kilobases (kb) upstream of the transcriptional start site of the human apolipoprotein B (apoB) gene, was sufficient to direct high-level expression of human apoB transgenes in mice. In this report, we expand our analysis of the distant apoB intestinal control region (ICR), by examining the function of segments in the vicinity of the 315 bp intestinal enhancer (315 IE). DNaseI hypersensitivity (DH) studies of a 4.8 kb segment from the ICR revealed three new DH sites, in addition to the previously described DH1 region present within the 315 IE. DH2 mapped to a 485 bp segment (485 IE) immediately upstream of the 315 IE that exhibited strong intestinal enhancer activity in transient transfection experiments with intestine-derived CaCo-2 cells. Within the DH2 region, an HNF-4/ARP-1 binding site was demonstrated by gel retardation experiments. A 1.8 kb segment incorporating the 485 IE was capable of driving expression of human apoB transgenes in the intestines of mice. Additionally, a third component of the apoB ICR was found about 1.2 kb downstream of the 315 IE, within a 1031 bp segment (1031 IE) that also harbored two DH sites, DH3 and DH4. This segment did not display enhancer activity but was capable of driving transgene expression in the intestine. The three components of the ICR displayed a similar pattern of apoB mRNA expression along the horizontal axis of the intestine. The previously characterized in vivo liver-specific elements of the apoB gene, namely, the second intron enhancer and the 5‘ upstream liver enhancer, did not play a role in intestinal expression of apoB transgenes in mice.