Wölwer Christina B, Pase Luke B, Pearson Helen B, Gödde Nathan J, Lackovic Kurt, Huang David C S, Russell Sarah M, Humbert Patrick O
Cell Cycle and Cancer Genetics, Peter MacCallum Cancer Centre, Melbourne, Australia.
Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia.
PLoS One. 2015 Nov 16;10(11):e0142655. doi: 10.1371/journal.pone.0142655. eCollection 2015.
Erythroid enucleation is critical for terminal differentiation of red blood cells, and involves extrusion of the nucleus by orthochromatic erythroblasts to produce reticulocytes. Due to the difficulty of synchronizing erythroblasts, the molecular mechanisms underlying the enucleation process remain poorly understood. To elucidate the cellular program governing enucleation, we utilized a novel chemical screening approach whereby orthochromatic cells primed for enucleation were enriched ex vivo and subjected to a functional drug screen using a 324 compound library consisting of structurally diverse, medicinally active and cell permeable drugs. Using this approach, we have confirmed the role of HDACs, proteasomal regulators and MAPK in erythroid enucleation and introduce a new role for Cyclin-dependent kinases, in particular CDK9, in this process. Importantly, we demonstrate that when coupled with imaging analysis, this approach provides a powerful means to identify and characterize rate limiting steps involved in the erythroid enucleation process.
红细胞去核对于红细胞的终末分化至关重要，涉及正成红细胞挤出细胞核以产生网织红细胞。由于同步化红细胞的困难，去核过程的分子机制仍知之甚少。为了阐明控制去核的细胞程序，我们采用了一种新型化学筛选方法，通过该方法，在体外富集准备去核的正成红细胞，并使用由结构多样、具有药用活性且可透过细胞的药物组成的324种化合物文库进行功能药物筛选。使用这种方法，我们证实了组蛋白去乙酰化酶、蛋白酶体调节剂和丝裂原活化蛋白激酶在红细胞去核中的作用，并在此过程中引入了细胞周期蛋白依赖性激酶，特别是CDK9的新作用。重要的是，我们证明，当与成像分析相结合时，这种方法提供了一种强大的手段来识别和表征红细胞去核过程中涉及的限速步骤。
