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REVIEW JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in Cell Cycle

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Leukemia (2004) 18, 189–218 & 2004 Nature Publishing Group All rights reserved 0887-6924/04 $25.00 www.nature.com/leu REVIEW JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in cell cycle progression and leukemogenesis LS Steelman1, SC Pohnert2, JG Shelton1, RA Franklin1,3, FE Bertrand1 and JA McCubrey1,3 1Department of Microbiology and Immunology, Brody School of Medicine at East Carolina University, Greenville, NC, USA; 2Department of Biochemistry, Brody School of Medicine at East Carolina University, Greenville, NC, USA; and 3Leo Jenkins Cancer Center, Brody School of Medicine at East Carolina University, Greenville, NC, USA The roles of the JAK/STAT, Raf/MEK/ERK and PI3K/Akt signal were known under different names including persisting-cell- transduction pathways and the BCR-ABL oncoprotein in stimulating-factor, Thy-1-inducing factor, multilineage hemato- leukemogenesis and their importance in the regulation of cell cycle progression and apoptosis are discussed in this review. poietic growth factor, histamine-producing cell-stimulating These pathways have evolved regulatory proteins, which serve factor, multicolony stimulating factor (CSF), CFU-stimulating to limit their proliferative and antiapoptotic effects. Small factor, eosinophil-CSF, megakaryocyte-CSF, erythroid-CSF, molecular weight cell membrane-permeable drugs that target burst-promoting activity, neutrophil–granulocyte-CSF and he- these pathways have been developed for leukemia therapy. One mopoietin-2, before it was cloned.21 Once it was cloned, it was such example is imatinib mesylate, which targets the BCR-ABL shown to have all these activities. The human IL-3 peptide kinase as well as a few structurally related kinases. This drug consists of 133 amino acids while the mouse peptide is 140 has proven to be effective in the treatment of CML patients. 21–24 However, leukemic cells have evolved mechanisms to become amino acids in length. The biological activity of the IL-3 22 resistant to this drug. A means to combat drug resistance is to molecule lies in residues 17–118, and is not dependent on the target other prominent signaling components involved in the glycosylation state of the protein.25 The half-life of the molecule pathway or to inhibit BCR-ABL by other mechanisms. Treat- is 40 min with clearance being mainly via re-absorption in renal ment of imatinib-resistant leukemia cells with drugs that target tubules.26 Ras (farnysyl transferase inhibitors) or with the protein IL-3 is primarily synthesized by T lymphocytes with a small destabilizer geldanamycin has proven to be a means to inhibit 1,2 the growth of resistant cells. This review will tie together three contribution from mast cells. IL-3 mRNA stability is regulated 27 important signal transduction pathways involved in the regula- by a rapamycin-sensitive pathway. This likely involves the tion of hematopoietic cell growth and indicate how their target of rapamycin (TOR) and its downstream target p70S6K. expression is dysregulated by the BCR-ABL oncoprotein. Frequent injections of mice with either IL-3 or inoculations of IL- Leukemia (2004) 18, 189–218. doi:10.1038/sj.leu.2403241 3-producing cells increase the number of myeloid, mast and Keywords: JAK; Ras; Raf; PI3K; BCR-ABL megakaryocytic cells.28,29 The IL-3 receptor (IL-3R) is a member of the cytokine receptor gene family and is comprised of a- and b-subunits.1 Both subunits contain extracellular domains common to the cytokine Hematopoietic cytokines and receptors 30–42 receptor family. The intracellular domains are more variable. The a subunit of the IL-3R is divided into three main Cytokines interact with cell-surface receptors initiating signaling domains: ligand-binding, transmembrane and intracellular cascades that promote cell growth and division, while inhibiting signaling domains. The ligand-binding domain has homology the pathways of apoptotic cell death.1–20 The JAK/STAT, Raf/ with other cytokine receptors (GM-CSF and IL-5); however, they MEK/ERK and PI3K/Akt signaling pathways are activated by a differ in their binding specificity. The intracellular signaling variety of cytokines that function to potentiate or inhibit domain of the a subunit lacks intrinsic kinase activity, but is hematopoiesis. These include IL-3, IL-7, SCF, G-CSF, type I S6K 30–32 required for activation of STAT5, Raf-1 and p70 . interferons and TGF-b. Signaling through IL-3/IL-3R interaction Transcriptional regulation of c-Fos and c-Jun is mediated has been widely studied as a prototype for how cytokine through the interaction of intracellular signaling domain with signaling activates JAK/STAT, Raf/MEK/ERK and PI3K/Akt path- 30 downstream targets. ways to modulate normal and abnormal hematopoiesis. The common b subunit is a member of the cytokine receptor IL-3 is a soluble highly glycosylated 26 kDa protein that superfamily, consisting of GM-CSF, IL-3 and IL-5 receptors. The belongs to the class of proteins called cytokines. The JAK/STAT, 33–36 b subunit also has no intrinsic kinase activity. The gene for Raf/MEK/ERK and PI3K/Akt signaling pathways elicited by IL-3 37 the b-subunit gene is located on mouse chromosome 15 and promote survival, growth and differentiation of pluripotent the a-chain gene is located on chromosome 14. The human b hematopoietic cells. A depiction of three pathways is presented chain is located on chromosome 22q13.1 and the a chain is diagrammatically in Figure 1. IL-3 was initially characterized by located on chromosome Xp22.3 (Table 1). The cytokine its ability to induce activity of 20-a-hydroxy-steroid-dehydro- receptor superfamily is characterized by a 200-amino-acid genase in spleen cells.21 It turns out that IL-3 was being studied extracellular domain consisting of two fibronectin b barrels. The by many different investigators, and the diverse bioactivities highly conserved consensus ‘WSXWS box’ is characteristic of the membrane proximal region of cytokine receptors.35,38 The Correspondence: JA McCubrey, Department of Microbiology and transduction of a signal by the cytokine receptor requires Immunology, Brody School of Medicine at East Carolina University, 600 Moye Blvd, Greenville, NC 27858, USA; Fax: þ 1 252 744 3104; conformational changes in the cytoplasmic domain of the b E-mail: [email protected] subunit. This conformational change promotes the binding and Received 29 August 2003; accepted 24 September 2003 activation of downstream targets including JAK, STAT and Roles of JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in leukemia LS Steelman et al 190 Figure 1 Overview of effects of IL-3 on JAK/STAT, Raf/MEK/ERK and PI3K/Akt signaling. The figure depicts some of the signal transduction pathways activated after ligation of the IL-3 receptor. Activation of the IL-3 receptor induces the activity of the JAK/STAT, Raf/MEK/ERK and PI3K/ Akt pathways, which results in cell proliferation and prevention of apoptosis. Activation of the JAK/STAT pathway can induce the expression of antiapoptotic genes such as Bcl-XL but it can also induce the expression of CIS and SOCS proteins, which serve as negative feedback regulators. IL- 3 can induce the Ras/Raf/MEK/ERK pathway, which results in phosphorylation of transcription factors that can promote cell proliferation. Akt can phosphorylate many targets (Bad, IkK, caspase-9, GSK3b, Foxo3), which have effects on apoptosis and cell cycle progression. Cytokines also induce the tyrosine phosphorylation of the Gab2 protein, which is a docking molecule. Gab2 associates with the Shp-2 phosphatase, Grb-2 and PI3PI3K. Gab2 can regulate PI3K and Ras activities. Cytokines also regulate the expression of phosphates such as Shp2 and SHIP1, which regulate these pathway. Dotted lines indicate potential interactions between the pathways. Solid lines indicate direct paths of signal transduction in the individual pathways. Table 1 Chromosomal locations of genes involved in signal like growth factor (IGF) signaling mediated by their receptors. transduction discussed in this review Gab2 serves as a docking site for PI3K and Shp2 and also connects with the Ras/Raf/MEK/ERK pathway via Grb-2. Gab-2 Gene Chromosomal Gene Chromosomal is also a target of BCR-ABL and will be discussed later. location location The cytoplasmic portion of the IL-3bc subunit is divided into two regions, which were identified by truncation mutants: the JAK1 1p32.3 Ha-Ras 11p15.5 35 JAK2 9p24 Ki-Ras 12p12.1 proximal and distal portions. The proximal portion of the JAK3 19p13.1 Raf-1 3p25 bc subunit interacts with and activates JAKs, SOCS, CIS, STATs, STAT1 2q32.2 A-Raf Xp11.2 c-Src, PI3K and Vav.1,35 In the area spanning the distal and STAT2 12q13.13 B-Raf 7q34 proximal regions of the bc chain are the binding sites for the Shc, STAT3 17q21 MEK1 15q21 Shp1 and Shp2 proteins. The distal portion the b subunit STAT4 2q32.2 ERK1 16p11.2 c mediates transcription of c-Fos and c-Jun via its ability to STAT5 17q11.2 ERK2 22q11.2 S6K 30,31 STAT6 17q13.3–q14.1 p85a PI3K 5q12–q13 promote activation of Raf-1 and p70 . At the very end of CIS 3p21.3 p85bPI3K 19q13.2 the membrane distal domain are regions that inhibit cell growth SOCS1 16p13 p110a PI3K 3q16.3 but promote viability.39 SOCS2 12q p110b 3q22.3 SOCS3 17q25.3 p110 PI3Ka˜ 7q22 SOCS4 18q22.2 p110 PI3Ka¨ 1p36.2 Roles of hematopoietic cytokine receptors in hematopoietic SOCS5 2p21 PTEN 10q23 Gab2 11q13.4 SHIP1 2q36–q37.1, 2 disease and neoplasia Grb2 17q24–q25 SHIP2 11q23 SHC 1q21 BCR 22q11.23 The A/J strain of mice is nonresponsive to IL-3 and contains a SHP-2 12q24 ABL 9q34.1 naturally occurring mutation in the IL-3Ra-chain gene.40 Knockout mice have been created that lack either the IL-3 or IL-3R genes.41,42 Mice lacking IL-3 or IL-3R or both have a similar phenotype.
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  • A-Raf and Raf-1 Work Together to Influence Transient ERK Phosphorylation and Gl/S Cell Cycle Progression

    A-Raf and Raf-1 Work Together to Influence Transient ERK Phosphorylation and Gl/S Cell Cycle Progression

    Oncogene (2005) 24, 5207–5217 & 2005 Nature Publishing Group All rights reserved 0950-9232/05 $30.00 www.nature.com/onc A-Raf and Raf-1 work together to influence transient ERK phosphorylation and Gl/S cell cycle progression Kathryn Mercer1, Susan Giblett1, Anthony Oakden2, Jane Brown2, Richard Marais3 and Catrin Pritchard*,1 1Department of Biochemistry, University of Leicester, Adrian Building, University Road, Leicester LEI 7RH, UK; 2Division of Biomedical Services, University of Leicester, University Road, Leicester LEI 7RH, UK; 3Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK The Raf/MEK/ERK (extracellular regulated kinase) Kerkhoff and Rapp, 1997, 1998; Woods et al., 1997; signal transduction pathway controls the ability of cells Marshall, 1999; Squires et al., 2002). This function to respond to proliferative, apoptotic, migratory and is mediated by the ability of ERKs to translocate to the differentiation signals. We have investigated the combined nucleus where they are able to phosphorylate transcrip- contribution of A-Raf and Raf-1 isotypes to signalling tion factors, particularly the AP-1 complex that through this pathway by generating mice with knockout comprises various heterodimers of c-fos (c-fos, FosB, mutations of both A-raf and raf-1 genes. Double knockout Fra-1, Fra-2) and c-jun (c-Jun, JunB, JunD) family (DKO) mice have a more severe phenotype than single null members (Whitmarsh and Davis, 1996; Balmanno mutations of either gene, dying in embryogenesis at E10.5. and Cook, 1999; Cook et al., 1999; Shaulian and The DKO embryos show no changes in apoptosis, but Karin, 2002).
  • High-Intensity Raf Signals Convert Mitotic Cell Cycling Into Cellular Growth'

    High-Intensity Raf Signals Convert Mitotic Cell Cycling Into Cellular Growth'

    (CANCERRESEARCH58. 1636-1640. April 15. 19981 Advances in Brief High-intensity Raf Signals Convert Mitotic Cell Cycling into Cellular Growth' Eugen Kerkhoff and Ulf R. Rapp2 lnstitutfürmedizinischeStrahlenkundeund Zellforschung(MSZ). Universityof Wurzburg,97078 Wiirzburg, Germany Abstract oncogernc c-Raf-1-estrogen receptor fusion protein (c-Raf-1-BxB ERTM). Activation of the exogenous kinase induces high and sustained The selectionof NIH 3T3 cellsexpressinga hydroxytamoxifen-induci c-Raf signals and inhibits DNA synthesis and mitosis. Despite the ble c-Raf-1-estrogen receptor fusion protein (c-Raf-1-BxB-ER@) in the inhibition of cell cycle progression, cell growth remains unaffected, absence or presence of the inducer results in dramatic differences in the expression levels of the fusion protein. Hydroxytamoxifen-mediated con leading to cells with a DNA content of G1 cells and a cell size of stitutive activation of the Rafsignal favors the selection ofcells expressing G2-M-phase cells. Therefore, high-intensity Raf signals uncouple the low levels of c-R.af-1-BxB-ER@. Cells selected in the absence of hy regulation of cell growth from DNA replication and mitosis. droxytamoxifen express up to 20 tImes higher levels of the inducible Raf kinase. Activation ofthe oncogenic Rafkinase in cells expressing low levels Materials and Methods leads to a weak activation of the Raf/Mek/Erk cascade and the induction of S phase In confluentcells.The activationof cellsexpressinghigh levels Expression Vector. The pBabe-puro-BxB-ER@ vector was constructed by ofthe kinaseleadsto a strongpersIstentsignalandinhibitsDNAsynthesis releasing sequences encoding the c-Raf-l-BxB-ER@ protein from the BJ4- andmitOsisinproliferatingcells.TheinhibitionofDNAsynthesisandcell BxB-ER@ vector (7) by EcoRI digestion (2.2-kb fragment) and inserting them dIvision is presumably due to the elevated expression ofthe cyclin-depend into the unique EcoRI recognition site of the pBabe-puro vector (13).