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DNA Methylation Regulates 5-Lipoxygenase Promoter Activity

  • Johannes Uhl
  • Niko Klan
  • Matthias Rose
  • Karl-Dieter Entian
  • Oliver Werz
  • Dieter Steinhilber
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 525)

Abstract

5-Lipoxygenase (5-LO), the key enzyme in leukotriene biosynthesis, is expressed in a tissue- and cell-differentiation specific manner. The 5-LO core promoter required for basal promoter activity has an unique G + C-rich sequence which contains five tandem Spl consensus sequences [1]. Previously, we could show that the 5-lipoxygenase promoter is regulated by DNA methylation [2]. Treatment of the 5-LO negative cell lines U937 and HL-60TB with the demethylating agent 5-aza-2′-deoxycytidine (AdC) upregulated 5-LO activity and the expression of 5-LO primary transcripts and mature mRNA indicating that AdC stimulates 5-LO expression at the level of gene transcription. Analysis of the methylation status by methylation-sensitive sequencing of the 5-LO promoter revealed that the core promoter region was methylated in U937 and HL-60TB cells, whereas it was unmethylated in the 5-LO positive wild-type HL-60 cell line. In transient reporter gene assays, there was a dramatic reduction of 5-LO promoter activity when the reporter gene constructs were methylated in-vitro by Sss I methylase before transfection, indicating that 5-LO promoter activity is strongly inhibited when the promoter is methylated.

Keywords

Promoter Activity Methylation Status U937 Cell Reporter Gene Construct Core Promoter Region 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Johannes Uhl
  • Niko Klan
  • Matthias Rose
  • Karl-Dieter Entian
  • Oliver Werz
    • 1
  • Dieter Steinhilber
    • 1
  1. 1.Institute of Pharmaceutical ChemistryUniversity of FrankfurtFrankfurtGermany

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