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Human Molecular Genetics Advance Access originally published online on June 9, 2004
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Human Molecular Genetics, 2004, Vol. 13, No. 15 1563-1575
DOI: 10.1093/hmg/ddh173
Human Molecular Genetics, Vol. 13, No. 15 © Oxford University Press 2004; all rights reserved

Photoreceptor-specific nuclear receptor NR2E3 functions as a transcriptional activator in rod photoreceptors

Hong Cheng1,2, Hemant Khanna2, Edwin C.T. Oh1,2, David Hicks4, Kenneth P. Mitton5 and Anand Swaroop1,2,3,*

1Neuroscience Graduate Program, 2Department of Ophthalmology and Visual Sciences and 3Department of Human Genetics, University of Michigan, Ann Arbor, MI, USA, 4Laboratoire de Physiopathologie Cellulaire et Moléculaire de la Rétine, INSERM Université Louis Pasteur E9918 Centre Hospitalier Universitaire Régional, 67091 Strasbourg, France and 5Eye Research Institute, Oakland University, Rochester, MI, USA

Received March 18, 2004; Accepted May 27, 2004

NR2E3, a photoreceptor-specific orphan nuclear receptor, is believed to play a pivotal role in the differentiation of photoreceptors. Mutations in the human NR2E3 gene and its mouse ortholog are associated with enhanced S-cones and retinal degeneration. In order to gain insights into the NR2E3 function, we performed temporal and spatial expression analysis, yeast two-hybrid screening, promoter activity assays and co-immunoprecipitation studies. The Nr2e3 expression was localized preferentially to the rod, and not to the cone, photoreceptor nuclei in rodent retina. The yeast two-hybrid screening of a retinal cDNA library, using NR2E3 as the bait, identified another orphan nuclear receptor NR1D1 (Rev-erb{alpha}). The interaction of NR2E3 with NR1D1 was confirmed by glutathione S-transferase pulldown and co-immunoprecipitation experiments. In transient transfection studies using HEK 293 cells, both NR2E3 and NR1D1 activated the promoters of rod phototransduction genes synergistically with neural retina leucine zipper (NRL) and cone–rod homeobox (CRX). All four proteins, NR2E3, NR1D1, NRL and CRX, could be co-immunoprecipitated from the bovine retinal nuclear extract, suggesting their existence in a multi-protein transcriptional regulatory complex in vivo. Our results demonstrate that NR2E3 is involved in regulating the expression of rod photoreceptor-specific genes and support its proposed role in transcriptional regulatory network(s) during rod differentiation.

* To whom correspondence should be addressed at: Department of Ophthalmology and Visual Sciences, W.K. Kellogg Eye Center, University of Michigan, 1000 Wall Street, Ann Arbor, MI 48105, USA. Tel: +1 7347633731; Fax: +1 7346470228; Email: swaroop{at}umich.edu


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