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Transcription elongation factors are involved in programming hormone production in pituitary neuroendocrine GH4C1

Toshitsugu Fujita1, Isabelle Piuz, Werner Schlegel

  • 1Fondation pour Recherches Médicales, Medical Faculty of the University of Geneva, Geneva, Switzerland.

Molecular and Cellular Endocrinology
|January 26, 2010
PubMed

Insights

Negative transcription elongation factors, 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) sensitivity-inducing factor (DSIF) and negative elongation factor (NELF), regulate hormone expression. Knockdown studies revealed their crucial roles in prolactin and growth hormone production.

Area of Science:

  • Molecular Biology
  • Endocrinology
  • Gene Regulation

Background:

  • Transcription elongation is a critical regulatory step in eukaryotic gene expression.
  • Negative transcription elongation factors, DSIF and NELF, are known to pause RNA polymerase II.
  • The role of these factors in hormone production in neuroendocrine cells was previously unclear.

Purpose of the Study:

  • To investigate the role of DSIF and NELF in the regulation of hormone expression in rat pituitary GH4C1 cells.
  • To determine the specific effects of NELF and DSIF subunits on prolactin (PRL) and growth hormone (GH) expression.

Main Methods:

  • Stable knockdown of NELF-E (a NELF subunit) and Spt5 (a DSIF subunit) using gene silencing techniques.
  • Quantitative analysis of PRL and GH mRNA and protein levels.
  • Transient knockdown of NELF-E and assessment of PRL promoter activity using reporter gene assays.
  • Chromatin immunoprecipitation (ChIP) to assess direct NELF-gene interaction.

Main Results:

  • Stable knockdown of NELF-E or Spt5 increased PRL expression at both mRNA and protein levels.
  • Stable knockdown of Spt5 alone abolished GH expression.
  • Transient NELF-E knockdown enhanced PRL expression and PRL promoter activity.
  • ChIP assays did not show direct NELF binding to the PRL gene, suggesting indirect regulation.

Conclusions:

  • NELF and DSIF play significant roles in regulating PRL and GH expression in pituitary cells.
  • NELF appears to suppress PRL promoter activity indirectly.
  • Transcription regulation by NELF and DSIF is essential for hormone production and may influence neuroendocrine cell differentiation.

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