MAZ, a Myc-associated zinc finger protein, is essential for the ME1a1-mediated expression of the c-myc gene during neuroectodermal differentiation of P19 cells

To investigate whether MAZ (Myc-associated zinc finger protein) affects the expression of the c-myc gene during the retinoic acid-induced (RA-induced) neuroectodermal differentiation of P19 embryonal carcinoma (EC) cells, we introduced a CAT reporter construct, human c-myc promoter/CAT (pMyc2CAT), and a mutant CAT derivative that lacked an ME1a1 site (pMyc1CAT) into P19EC cells to monitor the promoter activity of the c-myc gene. The expression of CAT in pMyc2CAT-transformed cells declined fivefold after 24 h in the presence of RA, returned to the normal level within 48 h, and decreased again to below 20% of the normal level after 96 h. By contrast, the expression of CAT in pMyc1CAT-transformed cells did not return to the normal level after 48 h in the presence of RA. In addition, an electrophoretic mobility shift assay (EMSA) with ME1a1 DNA as probe demonstrated that the kinetics of the DNA-binding activity of MAZ were closely correlated with the changes in the expression of CAT from the c-myc promoter/CAT gene during the differentiation of P19EC cells. Taken together, these results suggest that MAZ plays a key role in the transient increase in the expression of the c-myc gene after 48 h of exposure to RA during the neuroectodermal differentiation of P19EC cells.