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Papers In Press, published online ahead of print September 4, 2002
Obstetrics and Gynecology, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario M5G 1X5
Corresponding Author: brown{at}mshri.on.ca
The aryl hydrocarbon receptor (AhR) is a ligand-activated member of the bHLH-PAS transcription factor family. This receptor has been shown to be important in various aspects of growth and development as demonstrated by AhR null mice. A yeast two-hybrid screen of a mouse embryonic day 11 library for AhR interacting proteins revealed Nedd8 as a candidate. The interaction was confirmed in a cell free system and in mammalian cells by co-immunoprecipitation; however, in vitro neddylation assays showed that Nedd8 does not covalently modify AhR. Transfection of Nedd8 into a cell line stably transfected with a dioxin response element linked to a CAT reporter gene demonstrated that Nedd8 amplified ligand-induced transcription. Deletion of the Gly-76 residue in the C-terminus of Nedd8 abolished this effect and prevented AhR:Nedd8 interaction. Nedd8 overexpression also resulted in accumulation of AhR protein in the nucleus, independent of exogenous ligand. These studies demonstrate the AhR interacts with Nedd8 and suggest that this interaction enhances the transcriptional activity of the receptor, perhaps involving increased nuclear accumulation or retention. Immunohistochemistry performed on E11.5 mouse sections indicated Nedd8 and AhR localize to overlapping areas in the heart and spinal ganglia, raising the possibility that this interaction may play a role in organogenesis.
J. Biol. Chem, 10.1074/jbc.M202413200
Submitted on March 12, 2002
Revised on September 4, 2002
Accepted on September 4, 2002
Interaction with NEDD8, a ubiquitin-like protein, enhances the transcriptional activity of th earyl hydrocarbon receptor
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