Genomic cloning, structure, and regulatory elements of the 1 alpha, 25(OH)2D3 down-regulated gene for cyclic AMP-dependent protein kinase inhibitor

Biochim Biophys Acta. 1999 Sep 3;1446(3):414-8. doi: 10.1016/s0167-4781(99)00101-3.

Abstract

The cyclic AMP-dependent protein kinase inhibitor (PKI) mRNA and protein are negatively and tissue-specifically regulated in the kidney by 1 alpha, 25(OH)2D3. A 17-kb PKI clone, isolated from a chick genomic library, revealed that the PKI gene consists of two exons separated by a 4.5-kb intron. A 411-bp upstream region (constituting 93 bp upstream and 318 bp downstream from the transcriptional start site) containing a putative negative VDRE (nVDRE) fused to the luciferase gene was used for transient transfections of primary cultures of chick kidney cells. Luciferase activity was significantly down-regulated in response to 1 alpha, 25(OH)2D3. This result suggests that the promoter region containing the putative nVDRE plays a pivotal role in the negative regulation of PKI gene transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Calcitriol / pharmacology*
  • Carrier Proteins / genetics*
  • Cells, Cultured
  • Chickens
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • Down-Regulation / drug effects
  • Gene Expression Regulation / drug effects
  • Intracellular Signaling Peptides and Proteins*
  • Kidney / metabolism
  • Luciferases / genetics
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Regulatory Sequences, Nucleic Acid
  • Transfection
  • beta-Galactosidase / genetics

Substances

  • Carrier Proteins
  • DNA, Complementary
  • Intracellular Signaling Peptides and Proteins
  • protein kinase modulator
  • Luciferases
  • beta-Galactosidase
  • Calcitriol

Associated data

  • GENBANK/AF139056