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Sequences that direct rat tyrosine hydroxylase gene expression.

B P Fung1, S O Yoon, D M Chikaraishi

  • 1Neuroscience Program, Tufts University School of Medicine, Boston, MA 02111.

Journal of Neurochemistry
|June 1, 1992
PubMed
Summary

A 70-bp region of the tyrosine hydroxylase (TH) gene, lacking a cyclic AMP response element (CRE), drives high reporter gene expression in neuronal cells. This region

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Gene Regulation

Background:

  • Neuroendocrine gene transcription is regulated by DNA binding sites, including the cyclic AMP response element (CRE).
  • The tyrosine hydroxylase (TH) gene is crucial for neuronal and chromaffin cell function.

Purpose of the Study:

  • To identify and characterize regulatory elements in the tyrosine hydroxylase (TH) gene promoter.
  • To investigate the role of a specific 70-bp region in conferring cell-specific expression and responsiveness to forskolin.

Main Methods:

  • Reporter gene assays (chloramphenicol acetyltransferase) were used to assess transcriptional activity.
  • Gel-shift assays were performed with nuclear extracts from TH-expressing cells.
  • Analysis of DNA binding sites including AP2, AP1, E2A/MyoD, and POU transcription factors.

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Main Results:

  • A 70-bp region (-229 to -160) of the TH gene, lacking a CRE, was sufficient for high reporter gene expression in neuronal cells.
  • This 70-bp region contains binding sites for AP2, AP1, and E2A/MyoD, which are involved in cell-specific complex formation.
  • The 70-bp region mediated a twofold transcriptional response to forskolin, despite lacking a canonical CRE.

Conclusions:

  • Promoter and enhancer elements are both critical for TH gene expression.
  • Specific transcription factors binding to the AP2, AP1, and E box sites in the 70-bp region are key regulators of TH expression.
  • The 70-bp region contributes to both basal and stimulated TH gene transcription in a cell-specific manner.