Synergistic activation of RLD-1 by agents triggering PKA and PKC dependent signalling

Ching Jang Huang, Dorothee Feltkamp, Stefan Nilsson, Jan Åke Gustafsson

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

RLD-1 and OR-1 are closely related orphan nuclear receptors that can be activated by certain oxysterols. To obtain cells stably expressing RLD-1 or OR-1, CHOK1 cells were successively transfected with a DGRE2-ALP reporter and GR- RLD-1 or GR-OR-1 chimeric constructs. The selected cell clones that showed low background activity of the reporter and maximum fold induction by 22R(OH)cholesterol were used for subsequent experiments. Treatment of the cells with PGE2, TPA, or 8-bromo-cAMP alone did not transactivate the reporter. However, the induction of the reporter by 22R(OH)cholesterol was markedly enhanced in the presence of PGE2, TPA, 8-bromo-cAMP, or forskolin in cells expressing GR-RLD-1. The enhancement was inhibited by H-89 and bisindolylmaleimide, both inhibitors of protein kinases. These results suggest that transactivation by ligand-activated RLD-1 may be further modulated/regulated through other signal transduction pathways involving phosphorylation catalyzed by protein kinases.

Original languageEnglish (US)
Pages (from-to)657-663
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume243
Issue number3
DOIs
StatePublished - Feb 24 1998

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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