Department of Pathology, Stanford University School of Medicine, Stanford, CA, 94305, USA.
Sun Yat-Sen University, School of Pharmaceutical Sciences, Guangzhou, 510006, China.
Sci Rep. 2019 Mar 20;9(1):4915. doi: 10.1038/s41598-019-41393-w.
The PBX1 homeodomain transcription factor is converted by t(1;19) chromosomal translocations in acute leukemia into the chimeric E2A-PBX1 oncoprotein. Fusion with E2A confers potent transcriptional activation and constitutive nuclear localization, bypassing the need for dimerization with protein partners that normally stabilize and regulate import of PBX1 into the nucleus, but the mechanisms underlying its oncogenic activation are incompletely defined. We demonstrate here that E2A-PBX1 self-associates through the PBX1 PBC-B domain of the chimeric protein to form higher-order oligomers in t(1;19) human leukemia cells, and that this property is required for oncogenic activity. Structural and functional studies indicate that self-association facilitates the binding of E2A-PBX1 to DNA. Mutants unable to self-associate are transformation defective, however their oncogenic activity is rescued by the synthetic oligomerization domain of FKBP, which confers conditional transformation properties on E2A-PBX1. In contrast to self-association, PBX1 protein domains that mediate interactions with HOX DNA-binding partners are dispensable. These studies suggest that oligomeric self-association may compensate for the inability of monomeric E2A-PBX1 to stably bind DNA and circumvents protein interactions that otherwise modulate PBX1 stability, nuclear localization, DNA binding, and transcriptional activity. The unique dependence on self-association for E2A-PBX1 oncogenic activity suggests potential approaches for mechanism-based targeted therapies.
PBX1 同源结构域转录因子通过急性白血病中的 t(1;19)染色体易位转化为嵌合 E2A-PBX1 癌蛋白。与 E2A 的融合赋予了强大的转录激活和组成型核定位,从而绕过了与通常稳定和调节 PBX1 核输入的蛋白伴侣二聚化的需求,但其致癌激活的机制尚未完全定义。我们在这里证明,E2A-PBX1 通过嵌合蛋白的 PBX1 PBC-B 结构域自我缔合,在 t(1;19)人类白血病细胞中形成更高阶的寡聚体,并且这种特性是致癌活性所必需的。结构和功能研究表明,自我缔合有助于 E2A-PBX1 与 DNA 的结合。不能自我缔合的突变体没有转化缺陷,然而,它们的致癌活性可以通过 FKBP 的合成寡聚化结构域挽救,该结构域赋予 E2A-PBX1 条件转化特性。与自我缔合相反,介导与 HOX DNA 结合伙伴相互作用的 PBX1 蛋白结构域是可有可无的。这些研究表明,寡聚体的自我缔合可能弥补了单体 E2A-PBX1 稳定结合 DNA 的能力的不足,并规避了否则会调节 PBX1 稳定性、核定位、DNA 结合和转录活性的蛋白相互作用。E2A-PBX1 致癌活性对自我缔合的独特依赖提示了针对机制的靶向治疗的潜在方法。