Cell Shape‐Based Chemical Screening Reveals an Epigenetic Network Mediated by Focal Adhesions
Cell Shape‐Based Chemical Screening Reveals an Epigenetic Network Mediated by Focal Adhesions
复制标题
基于细胞形状的化学筛选揭示了由局部粘附介导的表观遗传网络
DOI：
10.1111/febs.15840
复制
发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Okada Mariko
中科院分区：
文献类型：
--
作者：
Kanazawa Tomomi;Michida Hiroki;Uchino Yuki;Ishihara Akari;Zhang Suxiang;Tabata Sho;Suzuki Yutaka;Imamoto Akira;Okada Mariko
衔接蛋白CRK和CRKL参与多种信号传导途径，包括细胞粘附和哺乳动物细胞的命运调节。然而，CRK/CRKL在表观遗传调控中的分子功能在很大程度上仍然未知。在这里，我们开发了一个管道，使用高内容图像分析结合激酶和表观遗传调节剂的化学筛选来评估细胞形态。我们发现CRK/CRKL通过小鼠胚胎成纤维细胞的表观遗传改变来调节与细胞形态相关的基因调控网络。综合表观基因组和转录组分析显示，CRK/CRKL参与超级增强子活性和Cdt 1，Rin 1和Spp 1表达的上调，以调节细胞形态。对80种表观遗传抑制剂的筛选表明，组蛋白H3修饰剂、常染色质组蛋白甲基转移酶2和促分裂原和应激激活激酶1可能对CRK/CRKL介导的形态学变化很重要。综上所述，我们的研究结果表明，CRK/CRKL通过表观遗传修饰在基因调控网络中起着至关重要的作用。数据库染色质免疫沉淀测序和RNA测序数据分别以DRA 011080和DRA 011081登录号保存在日本DNA数据库中。
Adapter proteins CRK and CRKL participate in a variety of signaling pathways, including cell adhesion, and fate regulation of mammalian cells. However, the molecular functions of CRK/CRKL in epigenetic regulation remain largely unknown. Here, we developed a pipeline to evaluate cell morphology using high‐content image analysis combined with chemical screening of kinase and epigenetic modulators. We found that CRK/CRKL modulates gene regulatory networks associated with cell morphology through epigenetic alteration in mouse embryonic fibroblasts. Integrated epigenome and transcriptome analyses revealed that CRK/CRKL is involved in super‐enhancer activity and upregulation ofCdt1,Rin1, andSpp1expression for the regulation of cell morphology. Screening of a library of 80 epigenetic inhibitors showed that histone H3 modifiers, euchromatic histone methyltransferase 2 and mitogen‐ and stress‐activated kinase 1, may be important for CRK/CRKL‐mediated morphological changes. Taken together, our results indicate that CRK/CRKL plays a critical role in gene regulatory networks through epigenetic modification.DatabasesChromatin immunoprecipitation sequencing and RNA sequencing data were deposited in the DNA Data Bank of Japan under DRA011080 and DRA011081 accession numbers, respectively.