title: Phospholipase D2 (PLD2) shortens the time required for myeloid leukemic cell differentiation: mechanism of action.
abstract: Cell differentiation is compromised in acute leukemias. We report that mammalian target of rapamycin (mTOR) and S6 kinase (S6K) are highly expressed in the undifferentiated promyelomonocytic leukemic HL-60 cell line, whereas PLD2 expression is minimal. The expression ratio of PLD2 to mTOR (or to S6K) is gradually inverted upon in vitro induction of differentiation toward the neutrophilic phenotype. We present three ways that profoundly affect the kinetics of differentiation as follows: (i) simultaneous overexpression of mTOR (or S6K), (ii) silencing of mTOR via dsRNA-mediated interference or inhibition with rapamycin, and (iii) PLD2 overexpression. The last two methods shortened the time required for differentiation. By determining how PLD2 participates in cell differentiation, we found that PLD2 interacts with and activates the oncogene Fes/Fps, a protein-tyrosine kinase known to be involved in myeloid cell development. Fes activity is elevated with PLD2 overexpression, phosphatidic acid or phosphatidylinositol bisphosphate. Co-immunoprecipitation indicates a close PLD2-Fes physical interaction that is negated by a Fes-R483K mutant that incapacitates its Src homology 2 domain. All these suggest for the first time the following mechanism: mTOR/S6K down-regulation→PLD2 overexpression→PLD2/Fes association→phosphatidic acid-led activation of Fes kinase→granulocytic differentiation. Differentiation shortening could have a clinical impact on reducing the time of return to normalcy of the white cell counts after chemotherapy in patients with acute promyelocytic leukemia.
authors.affiliation: Department of Biochemistry and Molecular Biology, Wright State University School Medicine, Dayton, Ohio, 45435.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Di Fulvio
authors.affiliation: Department of Biochemistry and Molecular Biology, Wright State University School Medicine, Dayton, Ohio, 45435.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Frondorf
authors.affiliation: Department of Biochemistry and Molecular Biology, Wright State University School Medicine, Dayton, Ohio, 45435.
authors.forename: Karen M
authors.initials: KM
authors.lastname: Henkels
authors.affiliation: Department of Pharmacology and Toxicology, Wright State University School Medicine, Dayton, Ohio 45435.
authors.forename: William C
authors.initials: WC
authors.lastname: Grunwald
authors.affiliation: Department of Pharmacology and Toxicology, Wright State University School Medicine, Dayton, Ohio 45435.
authors.forename: David
authors.initials: D
authors.lastname: Cool
authors.affiliation: Department of Biochemistry and Molecular Biology, Wright State University School Medicine, Dayton, Ohio, 45435. Electronic address: julian.cambronero@wright.edu.
authors.forename: Julian
authors.initials: J
authors.lastname: Gomez-Cambronero
mesh_terms: D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D004789:Enzyme Activation / Q000187:drug effects / Q000235:genetics; D020868:Gene Silencing; D018922:HL-60 Cells; D006801:Humans; D007700:Kinetics; D007951:Leukemia, Myeloid / Q000473:pathology*; D008969:Molecular Sequence Data; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D051578:Proto-Oncogene Proteins c-fes / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D019893:Ribosomal Protein S6 Kinases / Q000235:genetics / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics; D018909:src Homology Domains
chemical_list: D012333:RNA, Messenger; C496645:FES protein, human; D051578:Proto-Oncogene Proteins c-fes; D019893:Ribosomal Protein S6 Kinases; D058570:TOR Serine-Threonine Kinases; C105606:phospholipase D2; D010739:Phospholipase D
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