Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon Hong Kong, China.
Protein Cell. 2010 Oct;1(10):927-34. doi: 10.1007/s13238-010-0114-y. Epub 2010 Nov 9.
Aberrant chromosomal fusion of the Ewing's sarcoma oncogene (EWS) to several different cellular partners produces the Ewing's family of oncoproteins (EWS-fusion-proteins, EFPs) and associated tumors (EFTs). EFPs are potent transcriptional activators, dependent on the N-terminal region of EWS (the EWS-activation-domain, EAD) and this function is thought to be central to EFT oncogenesis and maintenance. Thus EFPs are promising therapeutic targets, but detailed molecular studies will be pivotal for exploring this potential. Such studies have so far largely been restricted to intact mammalian cells while recent evidence has indicated that a mammalian cell-free transcription system may not support bona fide EAD function. Therefore, the lack of manipulatable assays for the EAD presents a significant barrier to progress. Using Xenopus laevis oocytes we describe a plasmid-based micro-injection assay that supports efficient, bona fide EAD transcriptional activity and hence provides a new vehicle for molecular dissection of the EAD.
Ewing 肉瘤致癌基因 (EWS) 与几个不同的细胞伙伴发生异常染色体融合，产生 Ewing 家族的癌蛋白（EWS-融合蛋白，EFPs）和相关肿瘤（EFTs）。EFPs 是有效的转录激活剂，依赖于 EWS 的 N 端区域（EWS-激活结构域，EAD），并且该功能被认为是 EFT 致癌和维持的核心。因此，EFPs 是很有前途的治疗靶点，但详细的分子研究对于探索这种潜力至关重要。到目前为止，这些研究主要局限于完整的哺乳动物细胞，而最近的证据表明，哺乳动物无细胞转录系统可能不支持真正的 EAD 功能。因此，缺乏可操作的 EAD 测定方法是进展的重大障碍。我们使用非洲爪蟾卵母细胞描述了一种基于质粒的微注射测定法，该测定法支持有效的、真正的 EAD 转录活性，因此为 EAD 的分子剖析提供了新的工具。
