No Relevance of NF-κB in the Transcriptional Regulation of Human Nanog Gene in Embryonic Carcinoma Cells
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Embryonic stem (ES) cells can self-renew maintaining the undifferentiated state. Self renewal requires many factors such as Oct4, Sox2, FoxD3, and Nanog. NF-κB is a transcription factor involved in many biological activities. Expression and activity of NF-kB increase upon differentiation of ES cells. Reportedly, Nanog protein directly binds to NF-κB protein and inhibits its activity in ES cells. Here, we found a potential binding site of NF-κB in the human Nanog promoter and postulated that NF-κB protein may regulate expression of the Nanog gene. We used human embryonic carcinoma (EC) cells as a model system of ES cells and confirmed decrease of Nanog and increase of NF-κB upon differentiation induced by retinoic acid. Although deletion analysis on the DNA fragment including NF-κB binding site suggested involvement of NF-κB in the negative regulation of the promoter, site-directed mutation of NF-κB binding site had no effect on the Nanog promoter activity. Furthermore, no direct association of NF-κB with the Nanog promoter was detected during differentiation. Therefore, we conclude that NF-κB protein may not be involved in transcriptional regulation of Nanog gene expression in EC cells and possibly in ES cells.
Abstract
Embryonic stem (ES) cells can self-renew maintaining the undifferentiated state. Self renewal requires many factors such as Oct4, Sox2, FoxD3, and Nanog. NF-κB is a transcription factor involved in many biological activities. Expression and activity of NF-kB increase upon differentiation of ES cells. Reportedly, Nanog protein directly binds to NF-κB protein and inhibits its activity in ES cells. Here, we found a potential binding site of NF-κB in the human Nanog promoter and postulated that NF-κB protein may regulate expression of the Nanog gene. We used human embryonic carcinoma (EC) cells as a model system of ES cells and confirmed decrease of Nanog and increase of NF-κB upon differentiation induced by retinoic acid. Although deletion analysis on the DNA fragment including NF-κB binding site suggested involvement of NF-κB in the negative regulation of the promoter, site-directed mutation of NF-κB binding site had no effect on the Nanog promoter activity. Furthermore, no direct association of NF-κB with the Nanog promoter was detected during differentiation. Therefore, we conclude that NF-κB protein may not be involved in transcriptional regulation of Nanog gene expression in EC cells and possibly in ES cells.
참고문헌 (20)
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