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Physiol. Genomics 34: 277-284, 2008. First published June 3, 2008; doi:10.1152/physiolgenomics.90236.2008
1094-8341/08 $8.00
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Received 30 April 2008; accepted in final form 30 May 2008.
Physiological Genomics 34:277-284 (2008)
1094-8341/08 $8.00 © 2008 American Physiological Society

SOX17 directly activates Zfp202 transcription during in vitro endoderm differentiation

Ethan S. Patterson1,2, Russell C. Addis1,2, Michael J. Shamblott1 and John D. Gearhart1,2

1 Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland
2 McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland

SOX17 is a SRY-related high-mobility group (HMG) box transcription factor that is necessary for endoderm formation in multiple species. Despite its essential function during endoderm formation and differentiation, few direct targets of SOX17 are known. To identify targets of SOX17, we isolated SOX17 binding sites with a chromatin immunoprecipitation (ChIP)-cloning screen. SOX17-ChIP identified zinc finger protein 202 (Zfp202) as a direct target of SOX17 during endoderm differentiation of F9 embryonal carcinoma cells. A sequence in the first intron of Zfp202 activated transcription in differentiated F9 cells, and overexpression of Sox17 increased the transcriptional activity of this sequence. SOX17 binds to a site within this sequence in electrophoretic mobility shift assays, and mutation of this site decreases the transcriptional activation. Zfp202 is induced concomitantly with Sox17 during endoderm differentiation of F9 cells. We also show that ZFP202 represses Hnf4a, which has been reported for the human ortholog ZNF202. Identifying targets of SOX17 will help to elucidate the molecular basis of endoderm differentiation and may provide a better understanding of the role of endoderm in patterning the other germ layers.

Sox17; chromatin immunoprecipitation; Hnf4a






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