title: A Chemical Screening Approach to Identify Novel Key Mediators of Erythroid Enucleation.
abstract: 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.
authors.affiliation: Cell Cycle and Cancer Genetics, Peter MacCallum Cancer Centre, Melbourne, Australia.|Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia.
authors.forename: Christina B
authors.initials: CB
authors.lastname: Wölwer
authors.affiliation: Cell Cycle and Cancer Genetics, Peter MacCallum Cancer Centre, Melbourne, Australia.|Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia.
authors.forename: Luke B
authors.initials: LB
authors.lastname: Pase
authors.affiliation: Cell Cycle and Cancer Genetics, Peter MacCallum Cancer Centre, Melbourne, Australia.|Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia.
authors.forename: Helen B
authors.initials: HB
authors.lastname: Pearson
authors.affiliation: Cell Cycle and Cancer Genetics, Peter MacCallum Cancer Centre, Melbourne, Australia.|Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia.
authors.forename: Nathan J
authors.initials: NJ
authors.lastname: Gödde
authors.affiliation: The Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia.|Department of Medical Biology, University of Melbourne, Melbourne, Australia.
authors.forename: Kurt
authors.initials: K
authors.lastname: Lackovic
authors.affiliation: The Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia.|Department of Medical Biology, University of Melbourne, Melbourne, Australia.
authors.forename: David C S
authors.initials: DC
authors.lastname: Huang
authors.affiliation: Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia.|Immune Signaling Laboratory, Peter MacCallum Cancer Centre, Melbourne, Australia.|Centre for Micro-Photonics, Faculty of Engineering and Industrial Sciences, Swinburne University of Technology, Hawthorn, Australia.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Russell
authors.affiliation: Cell Cycle and Cancer Genetics, Peter MacCallum Cancer Centre, Melbourne, Australia.|Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia.|Department of Pathology, University of Melbourne, Melbourne, Australia.|Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Patrick O
authors.initials: PO
authors.lastname: Humbert
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation; D042863:Cyclin-Dependent Kinase 9 / Q000378:metabolism; D004900:Erythroblasts / Q000187:drug effects* / Q000378:metabolism*; D004920:Erythropoiesis / Q000187:drug effects* / Q000502:physiology*; D005434:Flow Cytometry; D006655:Histone Deacetylases / Q000378:metabolism; D020935:MAP Kinase Signaling System; D051379:Mice; D008810:Mice, Inbred C57BL; D010641:Phenotype; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors / Q000737:chemistry; D012156:Reticulocytes / Q000166:cytology* / Q000502:physiology; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013678:Technology, Pharmaceutical / Q000379:methods*
chemical_list: D061988:Proteasome Inhibitors; C467972:Cdk9 protein, mouse; D042863:Cyclin-Dependent Kinase 9; D046988:Proteasome Endopeptidase Complex; D006655:Histone Deacetylases