April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Retiniods Regulate Rhodopsin Kinase Transcription via Receptor-Modulated Promoter Activity
Author Affiliations & Notes
  • S. C. Khani
    Ophthalmology, Schepens Eye Research Inst, Boston, Massachusetts
  • J. E. Young
    Ophthalmology, State University of New York, Buffalo, New York
  • Z. Khani-Oskouie
    Ophthalmology, Schepens Eye Research Inst, Boston, Massachusetts
  • G.-H. Peng
    Ophthalmology and Visual Sciences, Washington University School of Medicine, St Louis, Missouri
  • S. Chen
    Ophthalmology and Visual Sciences, Washington University School of Medicine, St Louis, Missouri
  • Footnotes
    Commercial Relationships  S.C. Khani, None; J.E. Young, None; Z. Khani-Oskouie, None; G.-H. Peng, None; S. Chen, None.
  • Footnotes
    Support  EY013600, EY016662, EY012543
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 1677. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      S. C. Khani, J. E. Young, Z. Khani-Oskouie, G.-H. Peng, S. Chen; Retiniods Regulate Rhodopsin Kinase Transcription via Receptor-Modulated Promoter Activity. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1677.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : Retinoids are powerful modulators of gene expression in a variety of cell types including photoreceptors where they are found in abundance. Many photoreceptor genes and promoters have retinoid response elements but relatively little is known about whether or not these sites are responsive to retinoid ligand signaling prior to and after differentiation. The purpose of the current study is to demonstrate the presence of active retinoid signaling pathways in the developing and mature photoreceptor cells using rhodopsin kinase (RK or GRK1) as a target gene model.The RK gene contains a uniquely compact promoter and plays a crucial role in photoreceptor recovery.

Methods: : Bioinformatic, gel retardation, yeast one-hybrid and chromatin immunoprecipitation studies were used to identify retinoid receptor binding sites on the RK promoter. RK expression levels, promoter activity and responsiveness to retinoids were analyzed using quantitative RT-PCR, immunostaining and immunoblotting of retinoblastoma cells or in RK-GFP transgenic mice. RK promoter fragments containing deletions were linked to luciferase reporter and tested by transient transfections for promoter activity and retinoid responsiveness.

Results: : Several putative retinoid response elements were found to interact with retinoid receptors primarily retinoid-X-receptor gamma (RXRγ). Chromatin immunoprecipitation further confirmed interaction of this retinoid receptor with Rk promoter in mouse retina. Exposure to retinoids stimulated expression of the endogenous Rk gene along with RK promoter-driven GFP or luciferase reporters not only in the retinoblastoma cells but also whole retinas of RK-GFP transgenic mice. The results of deletion/mutation analyses of the RK promoter suggested the presence of multiple retinoid responsive sites.

Conclusions: : In photoreceptors, retinoid receptor pathways modulate RK gene expression prior and after differentiation. Since RK promoter has been used in designing gene therapy for photoreceptor diseases, retinoids may prove useful in modulating RK-driven expression of therapeutic genes.

Keywords: photoreceptors • signal transduction: pharmacology/physiology • gene/expression 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×