UTILIZATION OF RECOMBINANT ADENOVIRUS AND DOMINANT-NEGATIVE MUTANTS TO CHARACTERIZE HEPATOCYTE NUCLEAR FACTOR 4-REGULATED APOLIPOPROTEIN AIAND CIII EXPRESSION

Using recombinant adenoviral vectors and a dominant negative mutant of HNF-4, we have examined the contribution of hepatocyte nuclear factor 4 (HNF-4) to endogenous apolipoprotein Al: and CIII mRNA expression. Overexpression of HNF-4 leads to a 7.4-fold increase in apolipoprotein CIII expression, while infection with the dominant negative mutant of HNF-4 reduces the level of apolipoprotein CIII mRNA by 80%, demonstra ting that endogenous HNF-4 is necessary for apolipoprotein CIII expres sion, Experiments using the hepatoma cell lines, HepG2 and Hep3B, indi cate that HNF-4 is also involved in the regulation of apolipoprotein A I expression in these lines, However, the effect of HNF-4 on apolipopr otein AI expression is much more dramatic in cell lines derived from i ntestinal epithelium, Infection of the intestinal-derived cell line IE C-6 with the HNF-4 adenovirus resulted in a greater than 20-fold incre ase in the level of apolipoprotein AI mRNA. These results indicate tha t HNF-4 does regulate apolipoprotein AI and CIII mRNA expression and s uggest that HNF-4 is critical for intestinal apolipoprotein AI express ion.