Functional characterization of the human phosphodiesterase 7A1 promoter

Biochem J. 2003 Aug 1;373(Pt 3):835-43. doi: 10.1042/BJ20021829.

Abstract

In this paper, the human phosphodiesterase 7A1 (h PDE7A1 ) promoter region was identified and functionally characterized. Transient transfection experiments indicated that a 2.9 kb fragment of the h PDE7A1 5'-flanking region, to position -2907, has strong promoter activity in Jurkat T-cells. Deletion analysis showed that the proximal region, up to position -988, contains major cis -regulatory elements of the h PDE7A1 promoter. This minimal promoter region contains a regulatory CpG island which is essential for promoter activity. The CpG island contains three potential cAMP-response-element-binding protein (CREB)-binding sites that, as judged by in vivo dimethyl sulphate (DMS) footprinting, are occupied in Jurkat T-cells. Moreover, over-expression of CREB results in increased promoter activity, but, on the other hand, promoter activity decreases when a dominant-negative form of CREB (KCREB) is over-expressed. In vivo DMS footprinting strongly indicates that other transcription factors, such Ets-2, nuclear factor of activated T-cells 1 (NFAT-1) and nuclear factor kappaB (NF-kappaB), might also contribute to the regulation of h PDE7A1 promoter. Finally, h PDE7A1 promoter was found to be induced by treatment with PMA, but not by treatment with dibutyryl cAMP or forskolin. These results provide insights into the factors and mechanisms that regulate expression of the h PDE7A gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3',5'-Cyclic-AMP Phosphodiesterases / chemistry
  • 3',5'-Cyclic-AMP Phosphodiesterases / genetics*
  • 3',5'-Cyclic-AMP Phosphodiesterases / metabolism
  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • Cloning, Molecular
  • Colforsin / pharmacology
  • CpG Islands
  • Cyclic AMP Response Element-Binding Protein / metabolism
  • Cyclic Nucleotide Phosphodiesterases, Type 7
  • DNA Footprinting
  • DNA, Complementary
  • Humans
  • Molecular Sequence Data
  • Promoter Regions, Genetic*
  • Tetradecanoylphorbol Acetate / pharmacology

Substances

  • Cyclic AMP Response Element-Binding Protein
  • DNA, Complementary
  • Colforsin
  • 3',5'-Cyclic-AMP Phosphodiesterases
  • Cyclic Nucleotide Phosphodiesterases, Type 7
  • PDE7A protein, human
  • Tetradecanoylphorbol Acetate