Identification of a novel c-Myc protein interactor, JPO2, with transforming activity in medulloblastoma cells

c-Myc oncogene activation is critical in the pathogenesis of a spectrum of human malignancies. The c-Myc NH2-terminal domain (MycNTD) is essential for cellular transformation, and mediates critical protein interactions that modulate c-Myc oncogenic properties. In Medulloblastoma, the most common malignant pediatric brain tumor, deregulated c-Myc expression is linked with poorer disease phenotypes and outcomes. The biological basis for these associations is, however, not well understood. To better understand mechanisms underlying Myc-mediated transformation of Medulloblastoma, we sought to identify novel MycNTD protein interactors from a Medulloblastoma cell line library using a unique two-hybrid system. We identified a novel MycNTD binding protein, JPO2, which shows nuclear colocalization with c-Myc, and interacts with c-Myc both in vitro and in mammalian cells. In Rat1a transformation assays, JPO2 potentiates c-Myc transforming activity, and can complement a transformation-defective Myc mutant. Immunohistochemical studies indicate tumor-specific JPO2 expression in human Medulloblastoma, and an association of JPO2 expression with metastatic Tumors. Significantly, JPO2 expression induces colony formation in UW228, a Medulloblastoma cell line, whereas RNAi-mediated JPO2 knockdown impairs colony formation in UW228, and in Myc-transformed UW228 cells. These data provide evidence for biochemical and functional interaction between c-Myc and JPO2 in Medulloblastoma transformation. JPO2 is closely related to JPO1, a Myc transcriptional target with transforming activity. As tumor-specific JPO1 expression in human and murine Medulloblastoma has also been reported; these collective observations suggest important functional links between the novel JPO protein family and c-Myc in Medulloblastoma transformation.