Cloning of a coproporphyrinogen oxidase promoter regulatory element binding protein

Shinichiro Takahashi, Kazumichi Furuyama, Akira Kobayashi, Shigeru Taketani, Hideo Harigae, Masayuki Yamamoto, Kazuhiko Igarashi, Takeshi Sasaki, Norio Hayashi

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Coproporphyrinogen oxidase [CPO] gene promoter regulatory element (CPRE) plays an important role in CPO gene regulation. To isolate a CPRE binding protein, we performed Southwestern screening of K562 cDNA expression library using CPRE as a probe and isolated a cDNA clone which encoded a novel protein, Klp1 (K562 cell-derived leucine-zipper-like protein 1). Klp1 mRNA was highly expressed in K562 cells, HeLa cells, and brain as a single transcript (1.4 kb). Gel mobility shift assays revealed that Klp1 specifically binds to CPRE. Computational analysis revealed that Klp1 has a leucine-zipper-like structure, a Leu-X-X-Leu-Leu motif, and a putative nuclear localization signal in the basic amino acid rich region. Transfection of the Klp1 expression vector into THP-1 cells resulted in transcriptional activation of a reporter construct containing CPRE. These results indicate that Klp1 is a DNA sequence-specific transcription factor that regulates gene expression of genes that contain CPRE in their regulatory region. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)596-602
Number of pages7
JournalBiochemical and biophysical research communications
Volume273
Issue number2
DOIs
Publication statusPublished - 2000 Jul 5

Keywords

  • CPO
  • CPRE
  • K562 cells
  • Klp1
  • Leu-X-X-Leu-Leu motif
  • Leucine-zipper-like protein
  • Southwestern screening

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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