Study of the role of retinoblastoma protein in terminal differentiation of murine erythroleukemia cells.
Hexamethylenebisacetamide-induced terminal differentiation of Friend virus-transformed murine erythroleukemia (MEL) cells can be inhibited by okadaic acid, an inhibitor of type 1 and type 2A protein phosphatases. The inhibition is shown to be correlated with prevention of dephosphorylation of retinoblastoma protein (pRB) in cells and bypass of G1 prolongation in the cell cycle. These results suggest that pRB-mediated G1 prolongation is necessary for MEL cells to commit to terminal differentiation. However, further experiments demonstrate that the simple cell cycle exit is not sufficient for commitment to terminal differentiation. Induction of dephosphorylation of pRB and subsequent G1 prolongation by forskolin does not lead MEL cells to differentiate. Additional pRB has been expressed in MEL cells by transfection with a neo-resistant plasmid containing RB cDNA under the control of a cytomegalovirus promoter. Exogenously expressed pRB is hyperphosphorylated in logarithmically growing MEL cells without any noticeable change in growth rate between the transfected cell line and the parental cell line. This result suggests that pRB in MEL cells is regulated by protein kinases and protein phosphatases and not by transcription.
Zhuo
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Laboratory of Neurochemistry, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892, USA.
Fan
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Huang
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Kaufman
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D000081:Acetamides / Q000494:pharmacology*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002453:Cell Cycle / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D002461:Cell Line, Transformed; D004988:Ethers, Cyclic / Q000494:pharmacology; D005622:Friend murine leukemia virus; D016193:G1 Phase; D015870:Gene Expression; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D016160:Retinoblastoma Protein / Q000096:biosynthesis / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000081:Acetamides; D000970:Antineoplastic Agents; D004988:Ethers, Cyclic; D016160:Retinoblastoma Protein; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; C014026:hexamethylene bisacetamide