ADENOVIRUS 5 E1A INDUCED-DIFFERENTIATION OF P19 EMBRYONAL CARCINOMA-CELLS REQUIRES BINDING TO P300

We transfected P19 embryonal carcinoma (EC) cells with genes encoding the adenovirus 5 E1A products. Expression of either the 12S or 13S transcripts yielded P19 cells either incapable of proliferating or able to proliferate but having lost the characteristics of the EC cell parent. The proliferating clones of E1A expressing P19 cells were incapable of differentiating in response to retinoic acid or dimethyl sulfoxide, no longer expressed the SSEA-1 surface antigen characteristic of EC cells, and did express cytokeratin 55, a marker of epithelial tissues. We used a number of 12S E1A constructs carrying deletions in the first exon and found that the effects on P19 cell growth and differentiated properties were lost with alterations affecting either the N terminal 25 amino acids or the CR1 region of the E1A protein. Both regions are required to bind the cellular p300 protein that we showed is present in P19 cells. We conclude that binding of E1A to the p300 protein in P19 cells results in the loss of EC cell characteristics.