Identification of RANBP16 and RANBP17 as novel interaction partners for the bHLH transcription factor E12.
Identification of RANBP16 and RANBP17 as novel interaction partners for the bHLH transcription factor E12.
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DOI：
10.1002/jcb.22689
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发表时间：
2010-09-01
影响因子：
4
通讯作者：
Smas, Cynthia M.
中科院分区：
文献类型：
--
作者：
Lee, Jun-Ho;Zhou, Shengli;Smas, Cynthia M.
关键词：
广泛表达的碱性螺旋-环-螺旋（bHLH）转录因子E12和E47是E2 A/TCF 3基因选择性剪接的产物，调节包括细胞生长、分化和发育在内的多种生物学过程。为了寻找E12的新蛋白质相互作用，我们利用E12的bHLH结构域作为诱饵在酵母双杂交筛选。酵母双杂交、哺乳动物双杂交和免疫共沉淀分析证明E12与RANBP 17（输入蛋白-β超家族的新成员）的特异性相互作用;这种相互作用映射到RANBP 17的CRM 1同源区。RANBP 17的异位表达导致E2 A/MyoD介导的E-box调节的荧光素酶报告基因的反式激活增加约3倍。相互作用和反式激活研究也揭示了RANBP 16/XPO 7的类似功能。此外，RANBP 16或RANBP 17的异位表达导致细胞周期蛋白依赖性激酶抑制剂p21 Waf 1/Cip 1（一种特征良好的E2 A靶基因）的内源性转录水平增加。总之，这些生物化学和功能数据揭示了RANBP 16和RANBP 17作为E2 A蛋白作用的新型调节剂。
The ubiquitously expressed basic helix-loop-helix (bHLH) transcription factors E12 and E47, products of alternative splicing of the E2A/TCF3 gene, regulate diverse biological processes including cell growth, differentiation and development. To search for novel protein interactions for E12, we utilized the bHLH domain of E12 as a bait in yeast two-hybrid screening. Yeast two-hybrid, mammalian two-hybrid, and co-immunoprecipitation analyses demonstrate specific interaction of E12 with RANBP17, a novel member of the importin-β superfamily; this interaction maps to the CRM1 homology region of RANBP17. Ectopic expression of RANBP17 leads to a ~3-fold increase in E2A/MyoD mediated transactivation of an E-box regulated luciferase reporter gene. Interaction and transactivation studies also revealed similar functions for RANBP16/XPO7. Furthermore, ectopic expression of either RANBP16 or RANBP17 resulted in increased level of endogenous transcript for the cyclin-dependent kinase inhibitor, p21Waf1/Cip1, a well-characterized E2A target gene. Together, these biochemical and functional data reveal RANBP16 and RANBP17 as novel regulators of E2A protein action.
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