Expression of rod-derived cone viability factor

Dual role of CRX in regulating promoter activity and cell-type specificity

Sophie Lambard, Sacha Reichman, Cynthia Ann Berlinicke, Marie Laure Niepon, Olivier Goureau, José Alain Sahel, Thierry Léveillard, Donald J Zack

Research output: Contribution to journalArticle

Abstract

Background: RdCVF and RdCVF2, encoded by the nucleoredoxin-like genes NXNL1 and NXNL2, are trophic factors with therapeutic potential that are involved in cone photoreceptor survival. Studying how their expression is regulated in the retina has implications for understanding both their activity and the mechanisms determining cell-type specificity within the retina. Methodology/Principal Findings: In order to define and characterize their promoters, a series of luciferase/GFP reporter constructs that contain various fragments of the 5′-upstream region of each gene, both murine and human, were tested in photoreceptor-like and non-photoreceptor cell lines and also in a biologically more relevant mouse retinal explant system. For NXNL1, 5′-deletion analysis identified the human -205/+57 bp and murine -351/+51 bp regions as having promoter activity. Moreover, in the retinal explants these constructs drove expression specifically to photoreceptor cells. For NXNL2, the human -393/+27 bp and murine -195/+70 bp regions were found to be sufficient for promoter activity. However, despite the fact that endogenous NXNL2 expression is photoreceptor-specific within the retina, neither of these DNA sequences nor larger upstream regions demonstrated photoreceptor-specific expression. Further analysis showed that a 79 bp NXNL2 positive regulatory sequence (-393 to 315 bp) combined with a 134 bp inactive minimal NXNL1 promoter fragment (-77 to +57 bp) was able to drive photoreceptor-specific expression, suggesting that the minimal NXNL1 fragment contains latent elements that encode cell-type specificity. Finally, based on bioinformatic analysis that suggested the importance of a CRX binding site within the minimal NXNL1 fragment, we found by mutation analysis that, depending on the context, the CRX site can play a dual role. Conclusions/Significance: The regulation of the Nucleoredoxin-like genes involves a CRX responsive element that can act as both as a positive regulator of promoter activity and as a modulator of cell-type specificity.

Original languageEnglish (US)
Article numbere13075
JournalPLoS One
Volume5
Issue number10
DOIs
StatePublished - 2010

Fingerprint

rods (retina)
Vertebrate Photoreceptor Cells
cones (retina)
photoreceptors
Retina
Cones
Genes
promoter regions
viability
retina
Retinal Cone Photoreceptor Cells
mice
Photoreceptor Cells
DNA sequences
cells
Bioinformatics
Computational Biology
Luciferases
explants
Modulators

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Expression of rod-derived cone viability factor : Dual role of CRX in regulating promoter activity and cell-type specificity. / Lambard, Sophie; Reichman, Sacha; Berlinicke, Cynthia Ann; Niepon, Marie Laure; Goureau, Olivier; Sahel, José Alain; Léveillard, Thierry; Zack, Donald J.

In: PLoS One, Vol. 5, No. 10, e13075, 2010.

Research output: Contribution to journalArticle

Lambard, Sophie ; Reichman, Sacha ; Berlinicke, Cynthia Ann ; Niepon, Marie Laure ; Goureau, Olivier ; Sahel, José Alain ; Léveillard, Thierry ; Zack, Donald J. / Expression of rod-derived cone viability factor : Dual role of CRX in regulating promoter activity and cell-type specificity. In: PLoS One. 2010 ; Vol. 5, No. 10.
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T2 - Dual role of CRX in regulating promoter activity and cell-type specificity

AU - Lambard, Sophie

AU - Reichman, Sacha

AU - Berlinicke, Cynthia Ann

AU - Niepon, Marie Laure

AU - Goureau, Olivier

AU - Sahel, José Alain

AU - Léveillard, Thierry

AU - Zack, Donald J

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