Regulation of human tyrosine hydroxylase gene by neuron-restrictive silencer factor

Biochem Biophys Res Commun. 2006 Jul 28;346(2):426-35. doi: 10.1016/j.bbrc.2006.05.142. Epub 2006 Jun 2.

Abstract

Tyrosine hydroxylase (TH), the biosynthetic enzyme of catecholamine, is synthesized specifically in catecholaminergic neurons. Thus, it is possible that neuronal cell type-specific expression of this gene is coordinately regulated. One of the neuron-specific transcription regulators, neuron-restrictive silencer factor (NRSF)/repressor element 1 (RE1) silencing transcription factor (REST), represses the expression of neuronal genes in non-neuronal cells. To elucidate the molecular mechanisms that control catecholaminergic neuronal expression of human TH, we initially characterized the 5' regulatory region. Previous studies have shown that a 3174 bp fragment of the human TH promoter confers specific expression to the reporter gene in dopaminergic neuron-like cell lines. Within this 5' regulatory region, three putative neuron-restrictive silencer elements (NRSE)/RE1 were identified, which bound NRSF/REST in a sequence-specific manner, as confirmed using EMSA and ChIP assays. In transient transfection assays, deletion or mutation of NRSE/RE1 elements led to a 7-fold increase in activity of the 3.2 kb TH promoter in human neural stem cells (NSCs), but had no major effects on differentiated neuron-like cells. Suppression of NRSF/REST functions with either the histone deacetylase inhibitor, trichostatin, or DN-NRSF induced TH promoter activity. Our data strongly suggest that NRSF/REST functions as a repressor of TH transcription in NSCs via a mechanism dependent on the TH NRSE/RE1 sites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Differentiation
  • Cell Line
  • Enzyme Activation
  • Gene Expression Regulation
  • Histone Deacetylase Inhibitors
  • Humans
  • Mutation
  • Neurons / cytology
  • Neurons / metabolism
  • Promoter Regions, Genetic
  • Protein Binding
  • Regulatory Elements, Transcriptional
  • Repressor Proteins / genetics
  • Repressor Proteins / physiology*
  • Stem Cells / cytology
  • Stem Cells / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / physiology*
  • Transcriptional Activation
  • Tyrosine 3-Monooxygenase / biosynthesis*
  • Tyrosine 3-Monooxygenase / genetics

Substances

  • Histone Deacetylase Inhibitors
  • RE1-silencing transcription factor
  • Repressor Proteins
  • Transcription Factors
  • Tyrosine 3-Monooxygenase