Department of Pharmaceutical Sciences, University of Connecticut, 69 North Eagleville Rd, Storrs, CT 06029-3092, USA.
Molecules. 2020 Mar 25;25(7):1500. doi: 10.3390/molecules25071500.
While loss-of-function mutations in the gene have been implicated as a driving force for a variety of pediatric brain tumors, as well as pancreatic neuroendocrine tumors, the role of ATRX in gene regulation and oncogenic development is not well-characterized. The ADD domain of ATRX (ATRX) localizes the protein to chromatin by specifically binding to the histone H3 tail. This domain is also a primary region that is mutated in these cancers. The overall goal of our studies was to utilize a variety of techniques (experimental and computational) to probe the H3:ATRX protein-protein interaction (PPI). We developed two biochemical assays that can be utilized to study the interaction. These assays were utilized to experimentally validate and expand upon our previous computational results. We demonstrated that the three anchor points in the H3 tail (A1, K4, and K9) are all essential for high affinity binding and that disruption of more than one contact region will be required to develop a small molecule that disrupts the PPI. Our approach in this study could be applied to other domains of ATRX, as well as PPIs between other distinct proteins.
虽然基因中的功能丧失突变被认为是多种儿科脑肿瘤以及胰腺神经内分泌肿瘤的驱动因素，但 ATRX 在基因调控和致癌发展中的作用尚未得到很好的描述。ATRX 的 ADD 结构域（ATRX）通过特异性结合组蛋白 H3 尾部将蛋白质定位到染色质上。该结构域也是这些癌症中突变的主要区域。我们研究的总体目标是利用各种技术（实验和计算）来探测 H3：ATRX 蛋白质-蛋白质相互作用（PPI）。我们开发了两种可用于研究相互作用的生化测定法。这些测定法被用于实验验证和扩展我们之前的计算结果。我们证明了 H3 尾部的三个锚定点（A1、K4 和 K9）对于高亲和力结合都是必不可少的，并且需要破坏多个接触区域才能开发出破坏 PPI 的小分子。我们在这项研究中的方法可以应用于 ATRX 的其他结构域以及其他不同蛋白质之间的 PPI。
