@article{Baobing Zhao_Ganesan Keerthivasan_Yang Mei_Jing Yang_James McElherne_Piu Wong_John G. Doench_Gang Feng_David E. Root_Peng Ji_2014, place={Pavia, Italy}, title={Targeted shRNA screening identified critical roles of pleckstrin-2 in erythropoiesis}, volume={99}, url={https://haematologica.org/article/view/7079}, DOI={10.3324/haematol.2014.105809}, abstractNote={Differentiation of erythroblasts to mature red blood cells involves dynamic changes of the membrane and cytoskeleton networks that are not fully characterized. Using a mouse fetal liver erythroblast culture system and a targeted shRNA functional screening strategy, we identified a critical role of pleckstrin-2 in actin dynamics and protection of early stage terminal erythroblasts from oxidative damage. Knockdown of pleckstrin-2 in the early stage of terminal erythropoiesis disrupted the actin cytoskeleton and led to differentiation inhibition and apoptosis. This pro-survival and differentiation function of pleckstrin-2 was mediated through its interaction with cofilin, by preventing cofilin’s mitochondrial entry when the intracellular level of reactive oxygen species was higher in the early stage of terminal erythropoiesis. Treatment of the cells with a scavenger of reactive oxygen species rescued cofilin’s mitochondrial entry and differentiation inhibition induced by pleckstrin-2 knockdown. In contrast, pleckstrin-2 knockdown in late stage terminal erythroblasts had no effect on survival or differentiation but blocked enucleation due to disorganized actin cytoskeleton. Thus, our study identified a dual function of pleckstrin-2 in the early and late stages of terminal erythropoiesis through its regulations of actin dynamics and cofilin’s mitochondrial localization, which reflects intracellular level of reactive oxygen species in different developmental stages.}, number={7}, journal={Haematologica}, author={Baobing Zhao and Ganesan Keerthivasan and Yang Mei and Jing Yang and James McElherne and Piu Wong and John G. Doench and Gang Feng and David E. Root and Peng Ji}, year={2014}, month={Jul.}, pages={1157-1167} }