Regulation of the orphan nuclear receptor Nr2f2 by the DFNA15 deafness gene Pou4f3.
Hair cells are the mechanotransducing cells of the inner ear that are essential for hearing and balance. POU4F3--a POU-domain transcription factor selectively expressed by these cells--has been shown to be essential for hair cell differentiation and survival in mice and its mutation in humans underlies late-onset progressive hearing loss (DFNA15). The downstream targets of POU4F3 are required for hair cell differentiation and survival. We aimed to identify such targets in order to elucidate the molecular pathways involved in hair cell production and maintenance. The orphan thyroid nuclear receptor Nr2f2 was identified as a POU4F3 target using a subtractive hybridization strategy and EMSA analysis showed that POU4F3 binds to two sites in the Nr2f2 5' flanking region. These sites were shown to be required for POU4F3 activation as their mutation leads to a reduction in the response of an Nr2f2 5' flanking region reporter construct to POU4F3. Immunocytochemistry was carried out in the developing and adult inner ear in order to investigate the relevance of this interaction in hearing. NR2F2 expression in the postnatal mouse organ of Corti was shown to be detectable in all sensory epithelia examined and characterised. These data demonstrate that Nr2f2 is a direct target of POU4F3 in vitro and that this regulatory relationship may be relevant to hair cell development and survival.
UCL Ear Institute, University College London, London, United Kingdom.
Chrysostomos
C
Tornari
UCL Ear Institute, University College London, London, United Kingdom.
Emily R
ER
Towers
UCL Ear Institute, University College London, London, United Kingdom.
Jonathan E
JE
Gale
UCL Ear Institute, University College London, London, United Kingdom.
Sally J
SJ
Dawson
D000818:Animals; D051837:COUP Transcription Factor II / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival; D006198:Hair Cells, Auditory / Q000166:cytology / Q000378:metabolism*; D034381:Hearing Loss / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009154:Mutation; D051381:Rats; D017207:Rats, Sprague-Dawley; D050821:Transcription Factor Brn-3C / Q000235:genetics / Q000378:metabolism*
D051837:COUP Transcription Factor II; D018398:Homeodomain Proteins; D050821:Transcription Factor Brn-3C; C497587:Nr2f2 protein, mouse; C497588:Nr2f2 protein, rat; C111223:POU4F3 protein, human; C121290:Pou4f3 protein, mouse; C095569:NR2F2 protein, human