Functional interaction of Yaf2 with the central region of MycN.

MYCN is often amplified in advanced-stage neuroblastomas with the consequence of enhanced MycN protein expression. By employing the yeast two-hybrid system we found that Yaf2 binds to the central region of MycN. Binding was also seen in vitro and in vivo. Ectopically expressed Yaf2, like MycN, is localized in the nuclei of neuroblastoma cells. Endogenous Yaf2 is expressed in all three tested neuroblastoma cell lines, all of which also express MycN. Yaf2 was able to enhance MycN-mediated transactivation from an E-box promoter, deletion of the Yaf2 binding region in MycN abrogates this effect. Thus, the binding of Yaf2 to the central region of MycN is functional in mammalian cells.