Identification of two distinct transactivation domains in the pluripotency sustaining factor nanog

Identification of a putative transactivation domain in human Nanog

“…The expression of the fusion protein of Gal4 DNA binding domain and hNanog individual domain plasmid constructs was confirmed by Western blot analysis with the anti-Gal4 DNA binding domain monoclonal antibody ( Figure 1B). Western blot analysis revealed that the additional fragment with larger molecular weight than we predicted was observed in Gal4/N-terminal fusion protein as in murine Nanog (Pan and Pei, 2003). To examine the transactivational potentials of hNanog individual regions in endogenously Nanog-expressing cells, NCCIT cells were transiently cotransfected with each fusion protein expression plasmid construct and reporter vector, pG5luc, containing 5 copies of Gal4 DNA binding sites in the upstream of the minimal promoter.…”

“…To analyze the transcriptional activity of hNanog, we cloned full length hNanog isoform lacking Lys (aa168) residue, designated as hNanog 1a (Unpublished data), from hEC cells and the open reading frame was roughly divided into three regions as studied in murine Nanog (Pan and Pei, 2003): N-terminal (aa1-94), homeodomain (aa95-154), and C-terminal (aa155-304) regions based on the amino acid sequences of homeodomain and the individual domains were fused to the yeast transcription factor, Gal4 DNA binding domain (aa1-147) lacking the capability of transcriptional activation by itself ( Figure 1A). The expression of the fusion protein of Gal4 DNA binding domain and hNanog individual domain plasmid constructs was confirmed by Western blot analysis with the anti-Gal4 DNA binding domain monoclonal antibody ( Figure 1B).…”

“…hNanog 1a was roughly divided into three regions based on the position of the homeodomain, as reported in murine Nanog (Pan and Pei, 2003). N-terminal (aa1-94), homeodomain (aa95-154) and C-terminal (aa155-304) regions of hNanog 1a were amplified by PCR using the full-length hNanog 1a cDNA as a template.…”

“…C-terminal region to the homeodomain contains novel Trp (W) repeat motif of unknown Identification of a putative transactivation dom ain in hum an Nanog structure and function. They reported that both N-and C-terminal regions to the homeodomain in the murine Nanog have active transcriptional activities and that the C-terminal region unexpectedly seems to be more active than the N-terminal region (Pan and Pei, 2003). hNanog shares about 57% overall and 87% identity within the homeodomain to the murine Nanog.…”

“…Recently, it has been reported that two distinct transactivational domains were identified in murine Nanog (Pan and Pei, 2003). Murine Nanog was divided into three regions based on the position of homeodomain.…”