title: EWS represses cofilin 1 expression by inducing nuclear retention of cofilin 1 mRNA.
abstract: In Ewing's sarcoma family tumors (ESFTs), the proto-oncogene EWS that encodes an RNA-binding protein is fused by chromosomal translocation to the gene encoding one of the E-twenty six (ETS) family of transcription factors, most commonly friend leukemia virus integration 1 (FLI-1). Although EWS/FLI-1 chimeric proteins are necessary for carcinogenesis, additional events seem to be required for transformation to occur. We have previously reported that a protein product of an EWS mRNA target, whose expression is negatively regulated by EWS but not by EWS/FLI-1, contributes to ESFT development. However, the mechanism by which EWS represses protein expression remains to be elucidated. Here, we report that overexpression of full-length EWS repressed protein expression and induced nuclear retention of reporter mRNAs in a tethering assay. In contrast, when a mutant lacking the EWS C-terminal nuclear localization signal (classified as a PY-NLS) was expressed, reporter protein expression was upregulated, and the number of cells exporting reporter mRNA to the cytoplasm increased. EWS binds to the 3'-untranslated region in another mRNA target, cofilin 1 (CFL1), and negatively regulates the expression of CFL1. Overexpression of EWS induced nuclear retention of CFL1 mRNA. Furthermore, ESFT cell proliferation and metastatic potential were suppressed by small interfering RNA-mediated CFL1 knockdown. Together, our findings suggest that EWS induces nuclear retention of CFL1 mRNA, thereby suppressing expression of CFL1, and that CFL1 promotes development of ESFT. Targeting CFL1 might therefore provide another novel approach for treatment of this aggressive disease.
authors.affiliation: 1] Molecular Entomology Laboratory, RIKEN, Wako, Japan [2] Department of Pathophysiology, Dalian Medical University, Dalian, China.
authors.forename: L
authors.initials: L
authors.lastname: Huang
authors.affiliation: Molecular Entomology Laboratory, RIKEN, Wako, Japan.
authors.forename: I
authors.initials: I
authors.lastname: Kuwahara
authors.affiliation: 1] Molecular Entomology Laboratory, RIKEN, Wako, Japan [2] PRESTO, Japan Science and Technology Agency, Kawaguchi, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Matsumoto
mesh_terms: D020413:3' Untranslated Regions; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D002471:Cell Transformation, Neoplastic; D051338:Cofilin 1 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D019913:Nuclear Localization Signals / Q000235:genetics / Q000378:metabolism; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D034802:RNA-Binding Protein EWS / Q000235:genetics / Q000502:physiology*; D012512:Sarcoma, Ewing / Q000235:genetics / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D051338:Cofilin 1; C000715470:MAS1 protein, human; D019913:Nuclear Localization Signals; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D034802:RNA-Binding Protein EWS