In Vitro Activity of the EWS Oncogene Transcriptional Activation Domain

King Pan Ng Kim K. C. Li and Kevin A. W. Lee

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In Vitro Activity of the EWS Oncogene Transcriptional Activation Domain

机译：EWS癌基因转录激活域的体外活性

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Aberrant chromosomal fusion of the Ewings sarcoma oncogene (EWS) to several differentncellular partners gives rise to the Ewing’s family of oncogenic proteins [EWS fusion proteins (EFPs)]nand associated tumors (EFTs). EFPs are potent transcriptional activators dependent on the N-terminalnregion of EWS [the EWS activation domain (EAD)], and this function is thought to be central to EFTnoncogenesis and maintenance. Thus, EFPs are promising therapeutic targets, and detailed molecular studiesnof the EAD will be pivotal for exploring this potential. For many reasons, the molecular mechanism ofnEAD action is poorly understood and one major obstacle to progress is the lack of an in vitro transcriptionnassay. Using well-characterized EAD-dependent activators and soluble nuclear extracts, we have attemptednto recapitulate EAD transcriptional actvity in vitro. We report that while the EAD activates transcriptionnstrongly in vitro, the effect of EAD mutations is strikingly different from that observed in vivo. Ournresults therefore suggest that crude soluble extracts do not support bona fide EAD activity in vitro, andnwe discuss our findings in relation to future assay development and potential mechanisms of EAD action.

机译：Ewings肉瘤癌基因（EWS）与几种不同细胞伴侣的异常染色体融合产生了Ewing的致癌蛋白家族[EWSs融合蛋白（EFPs）] nand相关肿瘤（EFTs）。 EFP是依赖于EWS的N末端区域[EWS激活域（EAD）]的有效转录激活因子，该功能被认为对EFTnoncogenesis和维持至关重要。因此，EFP是有希望的治疗靶标，EAD的详细分子研究对于探索这种潜力至关重要。由于许多原因，人们对nEAD作用的分子机制了解甚少，并且缺乏进展的主要障碍是缺乏体外转录检测。使用特征充分的EAD依赖性激活剂和可溶性核提取物，我们试图在体外概括EAD转录活性。我们报告说，虽然EAD在体外强烈激活转录，但EAD突变的作用与体内观察到的显着不同。因此，我们的结果表明，粗制可溶性提取物在体外不支持真正的EAD活性，因此我们讨论了与未来测定开发和EAD作用的潜在机制有关的发现。

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《Biochemistry》 |2009年第13期|p.2849-2857|共9页
作者
King Pan Ng Kim K. C. Li and Kevin A. W. Lee;
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Department of Biology, The Hong Kong UniVersity of Science and Technology, Clear Water Bay,Kowloon, Hong Kong, SAR China;

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专利

1. Mutation in the DNA-binding domain of the EWS-Oct-4 oncogene results in dominant negative activity that interferes with EWS-Oct-4-mediated transactivation. [J] . Kim S, Lee J, Kim JY, International Journal of Cancer =: Journal International du Cancer . 2009,第10期

机译：EWS-OCT-4癌基因的DNA结合结构域中的突变导致显性的阴性活性干扰EWS-OCT-4介导的转移激活。
2. EWS/FLI1 oncogene activates caspase 3 transcription and triggers apoptosis in vivo. [J] . Sohn EJ, Li H, Reidy Cancer research: The official organ of the American Association for Cancer Research, Inc . 2010,第3期

机译：EWS / FLI1癌基因在体内激活caspase 3转录并触发细胞凋亡。
3. RGG-boxes of the EWS oncoprotein repress a range of transcriptional activation domains [J] . Alex D, Lee KAW Nucleic Acids Research . 2005,第4期

机译：EWS癌蛋白的RGG框可抑制一系列转录激活域
4. Near-infrared oligonucleotide duplex sensors for imaging rapidly activated transcription factors in vitro and in situ [C] . Alexei A. Bogdanov Jr., Valeriy G. Metelev, Toloo Taghian, Dynamics and Fluctuations in Biomedical Photonics XVI . 2019

机译：用于在体外和原位快速激活转录因子成像的近红外寡核苷酸双链体传感器
5. Molecular Recognition by the EWS Transcriptional Activation Domain. [D] . Ng, Sheung Chun. 2013

机译：通过EWS转录激活域进行分子识别。
6. EWS/FLI1 Oncogene Activates Caspase 3 Transcription and Triggers Apoptosis In Vivo [O] . Eun Jung Sohn, Hongjie Li, Karen Reidy, -1

机译：EWS / FLI1癌基因激活Caspase 3转录并在体内触发细胞凋亡
7. Phosphorylation of the EWS IQ domain regulates transcriptional activity of the EWS/ATF1 and EWS/FLI1 fusion proteins [O] . R J Olsen, S H Hinrichs 2001

机译：EWS IQ结构域的磷酸化调节EWS / ATF1和EWS / FLI1融合蛋白的转录活性

In Vitro Activity of the EWS Oncogene Transcriptional Activation Domain

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