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.
Huang
Annie
A
Cancer Research Program, Canada. annie.huang@sickkids.ca
Ho
Cynthia S W
CS
Ponzielli
Romina
R
Barsyte-Lovejoy
Dalia
D
Bouffet
Eric
E
Picard
Daniel
D
Hawkins
Cynthia E
CE
Penn
Linda Z
LZ
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D048708:Cell Growth Processes / Q000502:physiology; D002467:Cell Nucleus / Q000378:metabolism; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D008527:Medulloblastoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D009842:Oligopeptides; D010455:Peptides / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D012097:Repressor Proteins; D014158:Transcription, Genetic; D015854:Up-Regulation
C513450:CDCA7L protein, human; D002843:Chromatin; C489427:MYC protein, human; D009687:Nuclear Proteins; D009842:Oligopeptides; D010455:Peptides; D016271:Proto-Oncogene Proteins c-myc; D012097:Repressor Proteins; C085706:FLAG peptide