Abstract
Nanog is a transcription factor identified by its ability to maintain the self-renewal of ES cells in the absence of leukemia inhibitory factor (LIF). Nanog protein contains an N-terminal domain (ND), a DNA-binding homeobox domain (HD) and a C-terminal domain (CD). We previously reported that the CD in Nanog is a transcriptional activation domain essential for the in vivo function of Nanog. Here we demonstrated that the ND in Nanog is also functionally important. Deletion of the ND reduces the transcriptional activity of Nanog on either artificial reporters or native Nanog promoters. This truncated Nanog is also less effective in regulating the endogenous Nanog target genes. Furthermore, the ND truncation disrupted the ability of Nanog to maintain ES cell self-renewal as well. We found that the ND is not required for the nuclear localization of Nanog. These results suggest that the regulation of endogenous pluripotent genes such as oct3/4 and rex-1 is required for the in vivo function of Nanog.
Similar content being viewed by others
References
Boyer L A, Lee T I, Cole M F, et al. Core transcriptional regulatory circuitry in human embryonic stem cells. Cell, 2005, 122: 947–956
Pan G J, Li J, Zhou Y L, et al. A negative feedback loop of transcription factors that controls stem cell pluripotency and self-renewal. Faseb J. 2006, 20: 1730–1732
Niwa H, Burdon T, Chambers I, et al. Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. Genes Dev, 1998, 12: 2048–2060
Silva J, Chambers I, Pollard S, et al. Nanog promotes transfer of pluripotency after cell fusion. Nature, 2006, 441: 997–1001
Yu J, Vodyanik M A, Smuga-Otto K, et al. Induced pluripotent stem cell lines derived from human somatic cells. Science, 2007, 318: 1917–1920
Pan G J, Pei D Q. The stem cell pluripotency factor NANOG activates transcription with two unusually potent subdomains at its C terminus. J Biol Chem, 2005, 280: 1401–1407
Wang Z, Ma TH, Chi X K, et al. Aromatic residues in the C-terminal domain 2 are required for nanog to mediate LIF-independent self-renewal of mouse embryonic stem cells. J Biol Chem, 2008, 283: 4480–4489
Mullin N P, Yates A, Rowe A J, et al. The pluripotency rheostat Nanog functions as a dimer. Biochem J, 2008, 411: 227–231
Shi W J, Wang H, Pan G J, et al. Regulation of the pluripotency marker Rex-1 by Nanog and Sox2. J Biol Chem, 2006, 281: 23319–23325
Chambers I, Colby D, Robertson M, et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell, 2003, 113: 643–655
Mitsui K, Tokuzawa Y, Itoh H, et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell, 2003, 113: 631–642
Hart A H, Hartley L, Ibrahim M, et al. Identification, cloning and expression analysis of the pluripotency promoting Nanog genes in mouse and human. Dev Dyn, 2004, 230: 187–198
Hyslop L, Stojkovic M, Armstrong L, et al. Downregulation of NANOG induces differentiation of human embryonic stem cells to extraembryonic lineages. Stem Cells, 2005, 23: 1035–1043
Pan G J, Pei D Q. Identification of two distinct transactivation domains in the pluripotency sustaining factor nanog. Cell Res, 2003, 13: 499–502
Loh Y H, Wu Q, Chew J L, et al. The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nat Genetics, 2006, 38: 431–440
Niwa H, Masui S, Chambers I, et al. Phenotypic complementation establishes requirements for specific POU domain and generic transactivation function of Oct-3/4 in embryonic stem cells. Mol Cell Biol, 2002, 22: 1526–1536
Author information
Authors and Affiliations
Corresponding author
Additional information
Contributed equally to this work
Supported by the National Natural Science Foundation of China (Grant Nos. 30725012, 30630039 and 90813033), Knowledge Innovation Project of Chinese Academy of Sciences (Grant No. KSCX2-YW-R-48), Guangzhou Science and Technology Commission Foundation (Grant No. 2008A1-E4011), National Key Basic Research and Development Program of China (Grant Nos. 2006CB701504, 2006CB943600, 2007CB948002 and 2007CB947804), and Bei**g Forestry University Young Scientist Fund (Grant No. Blx2w8003).
About this article
Cite this article
Guo, Y., Zhang, J., Ye, L. et al. The N-terminal domain is a transcriptional activation domain required for Nanog to maintain ES cell self-renewal. Chin. Sci. Bull. 54, 3271–3277 (2009). https://doi.org/10.1007/s11434-009-0530-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11434-009-0530-7