THE OCTAMER MOTIF PRESENT IN THE REX-1 PROMOTER BINDS OCT-1 AND OCT-3EXPRESSED BY EC CELLS AND ES CELLS

Rex-1 is a zinc finger-containing gene that is expressed in embryonal carcinoma (EC) cells and embryonic stem (ES) cells. Upon differentiati on with retinoic acid (RA), transcription of the Rex-1 gene decreases rapidly. Analysis of the 5'-flanking region of the Rex-1 gene identifi ed a consensus motif for the octamer family of transcription factors t hat stimulates expression from the Rex-1 promoter. In this report, we utilized gel mobility shift analysis to examine the binding of transcr iption factors to the Rex-1 octamer motif. F9 EC cells, D3 ES cells, a nd human NT2/D1 EC cells each form at least two prominent DNA/protein complexes with the octamer motif. Supershift analysis identifies Oct-1 and Oct-3 in these complexes. When F9 EC cells are induced to differe ntiate by treatment with RA for 48 h, there is a complete loss of the DNA/protein complex containing Oct-3. In contrast, the other DNA/prote in complexes, including the DNA/protein complex containing Oct-1, can still be detected. These results provide support for a role of Oct-3 i n the transcription of the Rex-1 gene. (C) 1994 Academic Press, Inc.