Regulation of apo A-IV transcription by lipid in newborn swine is associated with a promoter DNA-binding protein

Song Lu, Ying Yao, Heng Wang, Songmei Meng, Xiangying Cheng, Dennis Black

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Dietary lipid acutely upregulates apolipoprotein (apo) A-IV expression by sevenfold at the pretranslational 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 h. 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.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume284
Issue number2 47-2
StatePublished - Feb 1 2003

Fingerprint

DNA-Binding Proteins
Triglycerides
Swine
Lipids
Diet
Repressor Proteins
Jejunum
Transcriptional Activation
Genes
Small Intestine
apolipoprotein A-IV
Up-Regulation

All Science Journal Classification (ASJC) codes

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

Cite this

Regulation of apo A-IV transcription by lipid in newborn swine is associated with a promoter DNA-binding protein. / Lu, Song; Yao, Ying; Wang, Heng; Meng, Songmei; Cheng, Xiangying; Black, Dennis.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 284, No. 2 47-2, 01.02.2003.

Research output: Contribution to journalArticle

@article{4c385117a2174311bd18b903fdae3bc3,
title = "Regulation of apo A-IV transcription by lipid in newborn swine is associated with a promoter DNA-binding protein",
abstract = "Dietary lipid acutely upregulates apolipoprotein (apo) A-IV expression by sevenfold at the pretranslational 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 h. 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.",
author = "Song Lu and Ying Yao and Heng Wang and Songmei Meng and Xiangying Cheng and Dennis Black",
year = "2003",
month = "2",
day = "1",
language = "English (US)",
volume = "284",
journal = "American Journal of Physiology",
issn = "1931-857X",
publisher = "American Physiological Society",
number = "2 47-2",

}

TY - JOUR

T1 - Regulation of apo A-IV transcription by lipid in newborn swine is associated with a promoter DNA-binding protein

AU - Lu, Song

AU - Yao, Ying

AU - Wang, Heng

AU - Meng, Songmei

AU - Cheng, Xiangying

AU - Black, Dennis

PY - 2003/2/1

Y1 - 2003/2/1

N2 - Dietary lipid acutely upregulates apolipoprotein (apo) A-IV expression by sevenfold at the pretranslational 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 h. 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.

AB - Dietary lipid acutely upregulates apolipoprotein (apo) A-IV expression by sevenfold at the pretranslational 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 h. 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.

UR - http://www.scopus.com/inward/record.url?scp=0037305414&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037305414&partnerID=8YFLogxK

M3 - Article

VL - 284

JO - American Journal of Physiology

JF - American Journal of Physiology

SN - 1931-857X

IS - 2 47-2

ER -