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Am J Physiol Gastrointest Liver Physiol (October 16, 2002). doi:10.1152/ajpgi.00391.2002
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Articles in PresS, published online ahead of print October 16, 2002
Am J Physiol Gastrointest Liver Physiol, 10.1152/ajpgi.00391.2002
Submitted on September 11, 2002
Accepted on October 12, 2002

Regulation of Apo A-IV Transcription by Dietary Lipid In Newborn Swine Is Associated with a Proximal Promoter DNA-Binding Protein

Song Lu1, Ying Yao1, Heng Wang1, Songmei Meng1, Xiangying Cheng1, and Dennis D Black1*

1 Children's Foundation Research Center for Memphis at Le Bonheur Children's Medical Center, Pediatrics Dept., University of Tennessee Health Science Center, Memphis, TN, USA

* To whom correspondence should be addressed. E-mail: dblack{at}utmem.edu.

Dietary lipid acutely up-regulates apo A-IV expression by 7-fold at the pre-translational level in neonatal swine jejunum. To determine the mechanism of this regulation, two-day-old female swine received intraduodenal infusions of low- and high-triacylglycerol (TG) isocaloric diets for 24 hrs. Nuclear runoff assay confirmed apo A-IV gene transcriptional regulation by the high-TG diet. Footprinting analysis using the swine apo A-IV proximal promoter sequence (+14 to -246 bp) demonstrated three regions protected by the low-TG extracts. Of these three motifs, only ACCTTC showed 100% homology to the human sequence and was further studied. EMSA was performed using probes containing wild-type (WT) and mutant (M) motifs. A shift was noted with the low-TG nuclear extracts with the WT probe, but not with the M probe. Excess unlabeled free WT probe competed out the shift, whereas the M probe did not. No significant shift occurred with either probe using high-TG extracts. These results suggest that a repressor protein binds to the ACCTTC motif and becomes unbound during lipid absorption, allowing transcriptional activation of the apo A-IV gene in newborn swine small intestine.




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