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J Biol Chem, Vol. 274, Issue 8, 4954-4961, February 19, 1999

The -700/-310 Fragment of the Apolipoprotein A-IV Gene Combined with the -890/-500 Apolipoprotein C-III Enhancer Is Sufficient to Direct a Pattern of Gene Expression Similar to That for the Endogenous Apolipoprotein A-IV Gene

Johanne Le Beyec, Valérie Chauffeton, Horng-Yuan Kan^¶, Pierre-Luc Janvier, Charlotte Cywiner-Golenzer, François-Patrick Chatelet, Athina Despina Kalopissis, Vassilis Zannis^¶, Jean Chambaz, Martine Pinçon-Raymond, and Philippe Cardot

From  U.505 INSERM and UPRESA CNRS 7079, 15 rue de l'Ecole de Médecine, 75006 Paris, France and the ^¶ Section of Molecular Genetics, Cardiovascular Institute, Boston University Medical Center, Boston, Massachusetts 02118-2394

Spatial gene expression in the intestine is mediated by specific regulatory sequences. The three genes of the apoA-I/C-III/A-IV cluster are expressed in the intestine following cephalocaudal and crypt-to-villus axes. Previous studies have shown that the -780/-520 enhancer region of the apoC-III gene directs the expression of the apoA-I gene in both small intestinal villi and crypts, implying that other unidentified elements are necessary for a normal intestinal pattern of apoA-I gene expression. In this study, we have characterized transgenic mice expressing the chloramphenicol acetyltransferase gene under the control of different regions of the apoC-III and apoA-IV promoters. We found that the -890/+24 apoC-III promoter directed the expression of the reporter gene in crypts and villi and did not follow a cephalocaudal gradient of expression. In contrast, the 700/+10 apoA-IV promoter linked to the 500/890 apoC-III enhancer directed the expression of the reporter gene in enterocytes with a pattern of expression similar to that of the endogenous apoA-IV gene. Furthermore, linkage of the 700/310 apoA-IV distal promoter region to the 890/+24 apoC-III promoter was sufficient to restore the appropriate pattern of intestinal expression of the reporter gene. These findings demonstrate that the 700/310 distal region of the apoA-IV promoter contains regulatory elements that, in combination with proximal promoter elements and the 500/890 enhancer, are necessary and sufficient to restrict apoC-III and apoA-IV gene expression to villus enterocytes of the small intestine along the cephalocaudal axis.

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