A tetracycline-repressible transactivator approach suggests a shorter half-life for tyrosine hydroxylase mRNA

David Foster, John R Strong, William W. Morgan

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Long-term increases in catecholamine release result in elevated levels of the mRNA for tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of these compounds. This increase is due, in part, to increased transcription. However, recent evidence suggests that increased stability of TH mRNA may also play a role. One of the problems in studying the stability of the TH message is the limitation of current methods for assessing transcript half-life. In this study the regulation of the expression of the rat TH gene was placed under the control of a tetracycline (Tet)-repressible transactivator (tTA). In the absence of doxycycline (Dox), an analogue of Tet, TH mRNA was synthesized. However, when Dox was present, transcription of TH message was essentially totally suppressed, and the resulting degradation of the TH mRNA provided an index of the half-life of this message. With this approach the computed half-life of TH mRNA was significantly shorter than that determined following actinomycin D administration. This effect was not due to some unique feature of the chimeric gene used to synthesize TH mRNA or to an untoward effect of the Tet analogue used to suppress TH transcription.

Original languageEnglish (US)
Pages (from-to)137-146
Number of pages10
JournalBrain Research Protocols
Volume7
Issue number2
DOIs
StatePublished - Jun 2001

Fingerprint

Trans-Activators
Tyrosine 3-Monooxygenase
Tetracycline
Half-Life
Messenger RNA
Doxycycline
Dactinomycin
Genes
Catecholamines

Keywords

  • Catecholamines
  • mRNA stability
  • Post-transcriptional regulation
  • Tyrosine hydroxylase
  • Tyrosine hydroxylase gene

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

A tetracycline-repressible transactivator approach suggests a shorter half-life for tyrosine hydroxylase mRNA. / Foster, David; Strong, John R; Morgan, William W.

In: Brain Research Protocols, Vol. 7, No. 2, 06.2001, p. 137-146.

Research output: Contribution to journalArticle

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