哺乳动物细胞中抗增生性肽的高通量屏幕鉴定关键转录因子家族
A High-Throughput Screen for Antiproliferative Peptides in Mammalian Cells Identifies Key Transcription Factor Families
影响因子:3.90000
分区:生物学2区 / 生化研究方法1区
发表日期:2024 Nov 15
作者:
Shane M Liila-Fogarty, Grace E Boyum, Claire L Schwabe, Gaelen T Hess
摘要
转录因子（TFS）是多种疾病的有前途的治疗靶标。 TFS通过参与多种特定蛋白质 - 蛋白质相互作用来执行其细胞作用。例如，某些TFS的同型或异二聚化控制DNA结合，而基础转录机械或染色质修饰符的TFS与组件之间的相互作用也可能很关键。从理论上讲，小分子可用于破坏TF功能所需的特定蛋白质 - 蛋白质界面，但实际上，很难识别具有必要特异性和功效的小分子，这可能是由于通常是TF功能构成的广泛蛋白质蛋白质界面所致。但是，与小分子相比，肽具有提供破坏此类界面所需的特异性和功效的潜力。在这里，我们确定了约15种肽，这些肽使用来自人类核局部蛋白质的80-Mer蛋白区域（肽）的库的高通量合并屏幕抑制了白血病细胞的增殖。抗增殖性肽富含已知参与特定TF二聚化的区域，包括基本的亮氨酸拉链（BZIP）域家族。这些BZIP结构域之一，来自TF JDP2的JDP2; BZIP_1是顶部的抗增殖肽，将K562细胞的增殖降低了2倍。 JDP2; BZIP_1抑制了AP-1转录活性和表达JDP2的过表达，这表明肽通过天然JDP2机制影响了增殖。出乎意料的是，鉴于BZIP结构域的强烈保护，带注释的二聚化结构域之外的残基对于肽的抗磷酸化效力至关重要。肽介导的抗增生作用引发了K562细胞的红细胞分化，并增加了多个细胞系模型的G0/G1细胞。我们还发现，包括JDP2； BZIP_1在内的许多抗增生性肽不需要核定位信号来起作用，这是在治疗应用中传递这些肽的潜在益处。
Abstract
Transcription factors (TFs) are a promising therapeutic target for a multitude of diseases. TFs perform their cellular roles by participating in multiple specific protein-protein interactions. For example, homo- or heterodimerization of some TFs controls DNA binding, while interactions between TFs and components of basal transcriptional machinery or chromatin modifiers can also be critical. While, in theory, small molecules could be used to disrupt specific protein-protein interfaces required for TF function, in practice, it is difficult to identify small molecules with the necessary specificity and efficacy, likely due to the extensive protein-protein interfaces that often underlie TF function. However, in contrast to small molecules, peptides have the potential to provide both the specificity and efficacy required to disrupt such interfaces. Here, we identified ∼15 peptides that inhibit the proliferation of leukemia cells using a high-throughput pooled screen of a library of 80-mer protein regions (peptides) derived from human nuclear-localized proteins. The antiproliferative peptides were enriched for regions known to be involved in specific TF dimerization, including the basic leucine zipper (bZIP) domain family. One of these bZIP domains, JDP2;bZIP_1, from the TF JDP2, was the top antiproliferative peptide, reducing the proliferation of K562 cells by 2-fold. JDP2;bZIP_1 inhibited AP-1 transcriptional activity and phenocopied JDP2 overexpression, suggesting that the peptide affected proliferation through a native JDP2 mechanism. Unexpectedly, given the strong conservation of the bZIP domain, residues outside of the annotated dimerization domain were critical for the peptide's antiproliferative potency. The peptide-mediated antiproliferative effect initiated erythrocyte differentiation in K562 cells and increased G0/G1 cells across multiple cell line models. We also found that many of the antiproliferative peptides identified in this study, including JDP2;bZIP_1, did not require a nuclear localization signal to function, a potential benefit for delivering these peptides in therapeutic applications.