Regulation of human CYP11B1 and CYP11B2 promoters by transposable elements and conserved cis elements

Steroids. 2012 Jan;77(1-2):100-9. doi: 10.1016/j.steroids.2011.10.010. Epub 2011 Oct 31.

Abstract

CYP11B1 and CYP11B2 responsible for the final steps of cortisol and aldosterone synthesis, respectively, are believed to be duplicate genes with distinctive promoters. Our sequence analysis uncovers that these two genes share great homology in the proximal upstream regions, but insertion of Alu and L1 elements drives promoters divergent. Each CYP11B promoter contains two Alu elements embedded in a truncated L1 element, breaking L1 into three disconnected fragments. Alu functions as an enhancer in both genes regardless of orientation and copy number. Insertion of Alu upstream of a SV40 promoter also elevates promoter activity. However, the effect of Alu on CYP11B1 is blocked by a second L1 element (CYP11B1-L1.2) inserted between the first one and the conserved proximal upstream region. Although CYP11B1-L1.2 is 5'-truncated and lacks a functional ORF, replacing it with a fluorescent gene demonstrates that the element can be transcribed from the CYP11B1 core promoter in an opposite direction and a smaller magnitude compared to CYP11B1. Deletion of CYP11B1-L1.2 greatly increases CYP11B1 promoter activity and restores the enhancing effect of Alu. The Ad5 and SF-1 binding elements conserved in the proximal core promoter play a role in basal expression of both genes. Mutation of the Ad5 site reduces promoter activity to the minimal level. ERRα is the transcription factor interacting with Ad5 during basal expression. The core promoters of both genes are also conserved in mouse and rat despite the fact that the sites corresponding to cre, Ad5, and SF-1 in rodent Cyp11b1 promoters deviate from consensus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenal Cortex / cytology
  • Adrenal Cortex / metabolism*
  • Aldosterone / biosynthesis
  • Alu Elements
  • Animals
  • Cells, Cultured
  • Conserved Sequence
  • Cytochrome P-450 CYP11B2 / chemistry
  • Cytochrome P-450 CYP11B2 / genetics*
  • Cytochrome P-450 CYP11B2 / metabolism
  • DNA Transposable Elements*
  • Enhancer Elements, Genetic
  • Humans
  • Hydrocortisone / biosynthesis
  • Long Interspersed Nucleotide Elements
  • Mice
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Plasmids
  • Promoter Regions, Genetic*
  • Rats
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Steroid 11-beta-Hydroxylase / chemistry
  • Steroid 11-beta-Hydroxylase / genetics*
  • Steroid 11-beta-Hydroxylase / metabolism
  • Transfection

Substances

  • DNA Transposable Elements
  • Aldosterone
  • Cytochrome P-450 CYP11B2
  • Steroid 11-beta-Hydroxylase
  • Hydrocortisone