Characterization of an upstream regulatory element of the human apolipoprotein E gene, and purification of its binding protein from the human placenta

Dae Woong Jo, Trond P. Leren, Zung Yoon Yang, Young Ho Chung, John M. Taylor, Young Ki Paik

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

We have previously demonstrated that expression of the human apolipoprotein (apo) E gene is controlled by multiple regulatory elements in the promoter [Paik et al. (1988) J. Biol. Chem. 263, 13340-13349; Chang et al. (1990) J. Biol. Chem. 265, 9496-9504]. To extend these studies, we have characterized an element in the apoE gene promoter that spans nucleotides -101 to -89, upstream regulatory element 3 (URE3). Transcription of promoter/marker gene constructs in vitro showed that URE3 modulates gene expression. Gel mobolity shift assays of URE3 using human placental nuclear extracts detected a specific binding protein whose activity can be modulated by micromolar amounts of divalent copper and zinc. Competitive binding and gel shift assays with mutant oligonucleotides revealed critical nucleotides within URE3 required for its specific nuclear protein-binding activity. Gel filtration and oligonucleotide affinity chromatography were employed to isolate a URE3-binding protein (URE3BP) from human placental nuclear extracts. Purified URE3BP appears to be a Mr = 300,000 protein that is composed of four equally-sized basic subunits of Mr = 67,000. These studies indicate that URE3 is an active regulatory component of the apoE gene

Original languageEnglish
Pages (from-to)915-922
Number of pages8
JournalJournal of Biochemistry
Volume117
Issue number4
DOIs
Publication statusPublished - 1995 Apr

Fingerprint

Apolipoproteins E
Placental Extracts
Placenta
Purification
Carrier Proteins
Genes
Oligonucleotides
Gels
Nucleotides
Gene Components
Assays
Competitive Binding
Nuclear Proteins
Affinity Chromatography
Affinity chromatography
Protein Binding
Gel Chromatography
Zinc
Copper
Transcription

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Jo, Dae Woong ; Leren, Trond P. ; Yang, Zung Yoon ; Chung, Young Ho ; Taylor, John M. ; Paik, Young Ki. / Characterization of an upstream regulatory element of the human apolipoprotein E gene, and purification of its binding protein from the human placenta. In: Journal of Biochemistry. 1995 ; Vol. 117, No. 4. pp. 915-922.
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Characterization of an upstream regulatory element of the human apolipoprotein E gene, and purification of its binding protein from the human placenta. / Jo, Dae Woong; Leren, Trond P.; Yang, Zung Yoon; Chung, Young Ho; Taylor, John M.; Paik, Young Ki.

In: Journal of Biochemistry, Vol. 117, No. 4, 04.1995, p. 915-922.

Research output: Contribution to journalArticle

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