Human 15-lipoxygenase gene promoter: Analysis and identification of DNA binding sites for IL-13-induced regulatory factors in monocytes

Uddhav Kelavkar, Susheng Wang, Angel Montero, James Murtagh, Ketan Shah, Kamal Badr

Research output: Contribution to journalArticle

Abstract

In order to study the transcriptional control of 15-LO expression, we have cloned and sequenced the human 15-LO promoter region. The 15-LO promoter is associated with a CpG island at the 5'-end of the gene, and sequence analysis reveals putative Sp1 and Ap2 binding site/s and absence of TATA or CAAT motifs. Transcription is initiated at one major site. Using deletion constructs, we have defined an active promoter region of 1056 bp. Gel-shift assays revealed that transcriptional factor(s) induced only in response to IL-13 treatment of human peripheral blood monocytes bind to the 15-LO promoter DNA. Two regions, DP1 (-140 to -92 bp) and DP2 (-353 to -304 bp) of the promoter were essential for transcription in HeLa cells and human peripheral monocytes. Hela nuclear extracts contained a specific nuclear factor(s) binding to 15-LO promoter DNA which are distinct from those derived from IL-13-treated human peripheral monocyte nuclear extracts. In addition, fluorescent in situ hybridization (FISH) results refined the previous localization of 15-LO to human chromosome 17p13.3.

Original languageEnglish (US)
Pages (from-to)173-182
Number of pages10
JournalMolecular Biology Reports
Volume25
Issue number3
DOIs
StatePublished - Jul 1998
Externally publishedYes

Keywords

  • DNA (promoter) binding sites
  • Gene localization
  • Gene promoter
  • HeLa cells
  • Interleukin
  • Monocytes
  • Regulatory factors

ASJC Scopus subject areas

  • Genetics
  • Cell Biology
  • Biochemistry

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