SEARCH:   Advanced

Prepublished online as a Blood First Edition Paper on August 7, 2003; DOI 10.1182/blood-2003-05-1479. This Article Full Text (PDF) All Versions of this Article: 2003-05-1479v1 102/12/3938    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Zhang, J. Articles by Lodish, H. F Search for Related Content PubMed PubMed Citation Articles by Zhang, J. Articles by Lodish, H. F Social Bookmarking                   What's this? Submitted May 9, 2003 Accepted July 21, 2003 Role of Ras signaling in erythroid differentiation of mouse fetal liver cells: functional analysis by a flow cytometry-based novel culture system Jing Zhang, Merav Socolovsky, Alec W Gross, and Harvey F Lodish* Whitehead Institute for Biomedical Research, Cambridge, MA, USA; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA * Corresponding author; email: lodish{at}wi.mit.edu. Ras signaling plays an important role in erythropoiesis. Its function has been extensively studied in erythroid and non-erythroid cell lines as well as in primary erythroblasts, but inconclusive results using conventional colony-forming unit-erythroid (CFU-E) assays have been obtained concerning the role of Ras signaling in erythroid differentiation. Here we describe a novel culture system that supports terminal fetal liver erythroblast proliferation and differentiation and that closely recapitulates erythroid development in vivo. Erythroid differentiation is monitored step-by-step and quantitatively by a flow cytometry analysis; this analysis distinguishes CD71 and TER119 double-stained erythroblasts into different stages of differentiation. To study the role of Ras signaling in erythroid differentiation, different H-ras proteins were expressed in CFU-E progenitors and early erythroblasts using a bicistronic retroviral system and their effects on CFU-E colony formation and erythroid differentiation were analyzed. Only oncogenic H-ras but not dominant negative H-ras reduced CFU-E colony formation. Analysis of infected erythroblasts in our newly-developed system showed that oncogenic H-ras blocks terminal erythroid differentiation but not through promoting apoptosis of terminally differentiated erythroid cells. Rather, oncogenic H-ras promotes abnormal proliferation of CFU-E progenitors and early erythroblasts and supports their Epo-independent growth. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: L. A. Steiner, Y. Maksimova, V. Schulz, C. Wong, D. Raha, M. C. Mahajan, S. M. Weissman, and P. G. Gallagher Chromatin Architecture and Transcription Factor Binding Regulate Expression of Erythrocyte Membrane Protein Genes Mol. Cell. Biol., October 15, 2009; 29(20): 5399 - 5412. [Abstract] [Full Text] [PDF] A. D. Wood, E. Chen, I. J. Donaldson, S. Hattangadi, K. A. Burke, M. A. Dawson, D. Miranda-Saavedra, H. F. Lodish, A. R. Green, and B. Gottgens ID1 promotes expansion and survival of primary erythroid cells and is a target of JAK2V617F-STAT5 signaling Blood, August 27, 2009; 114(9): 1820 - 1830. [Abstract] [Full Text] [PDF] F. Marques-Garcia, N. Ferrandiz, R. Fernandez-Alonso, L. Gonzalez-Cano, M. Herreros-Villanueva, M. Rosa-Garrido, B. Fernandez-Garcia, J. P. Vaque, M. M. Marques, M. E. Alonso, et al. p73 Plays a Role in Erythroid Differentiation through GATA1 Induction J. Biol. Chem., August 7, 2009; 284(32): 21139 - 21156. [Abstract] [Full Text] [PDF] H. Bhatia, J. L. Hallock, A. Dutta, S. Karkashon, L. S. Sterner, T. Miyazaki, A. Dean, and J. A. Little Short-chain fatty acid-mediated effects on erythropoiesis in primary definitive erythroid cells Blood, June 18, 2009; 113(25): 6440 - 6448. [Abstract] [Full Text] [PDF] C. Catela, D. Bilbao-Cortes, E. Slonimsky, P. Kratsios, N. Rosenthal, and P. te Welscher Multiple congenital malformations of Wolf-Hirschhorn syndrome are recapitulated in Fgfrl1 null mice Dis. Model. Mech., May 1, 2009; 2(5-6): 283 - 294. [Abstract] [Full Text] [PDF] G. Chan, D. Kalaitzidis, T. Usenko, J. L. Kutok, W. Yang, M. G. Mohi, and B. G. Neel Leukemogenic Ptpn11 causes fatal myeloproliferative disorder via cell-autonomous effects on multiple stages of hematopoiesis Blood, April 30, 2009; 113(18): 4414 - 4424. [Abstract] [Full Text] [PDF] T. C. Luis, F. Weerkamp, B. A. E. Naber, M. R. M. Baert, E. F. E. de Haas, T. Nikolic, S. Heuvelmans, R. R. De Krijger, J. J. M. van Dongen, and F. J. T. Staal Wnt3a deficiency irreversibly impairs hematopoietic stem cell self-renewal and leads to defects in progenitor cell differentiation Blood, January 15, 2009; 113(3): 546 - 554. [Abstract] [Full Text] [PDF] A. M. Pilon, M. O. Arcasoy, H. K. Dressman, S. E. Vayda, Y. D. Maksimova, J. I. Sangerman, P. G. Gallagher, and D. M. Bodine Failure of Terminal Erythroid Differentiation in EKLF-Deficient Mice Is Associated with Cell Cycle Perturbation and Reduced Expression of E2F2 Mol. Cell. Biol., December 15, 2008; 28(24): 7394 - 7401. [Abstract] [Full Text] [PDF] A. Elorza, B. Hyde, H. K. Mikkola, S. Collins, and O. S. Shirihai UCP2 Modulates Cell Proliferation through the MAPK/ERK Pathway during Erythropoiesis and Has No Effect on Heme Biosynthesis J. Biol. Chem., November 7, 2008; 283(45): 30461 - 30470. [Abstract] [Full Text] [PDF] M. A. Kerenyi, F. Grebien, H. Gehart, M. Schifrer, M. Artaker, B. Kovacic, H. Beug, R. Moriggl, and E. W. Mullner Stat5 regulates cellular iron uptake of erythroid cells via IRP-2 and TfR-1 Blood, November 1, 2008; 112(9): 3878 - 3888. [Abstract] [Full Text] [PDF] T. Yamashita, O. Ohneda, A. Sakiyama, F. Iwata, K. Ohneda, and Y. Fujii-Kuriyama The microenvironment for erythropoiesis is regulated by HIF-2{alpha} through VCAM-1 in endothelial cells Blood, August 15, 2008; 112(4): 1482 - 1492. [Abstract] [Full Text] [PDF] T. Sawado, J. Halow, H. Im, T. Ragoczy, E. H. Bresnick, M. A. Bender, and M. Groudine H3 K79 dimethylation marks developmental activation of the {beta}-globin gene but is reduced upon LCR-mediated high-level transcription Blood, July 15, 2008; 112(2): 406 - 414. [Abstract] [Full Text] [PDF] F. Lohmann and J. J. Bieker Activation of Eklf expression during hematopoiesis by Gata2 and Smad5 prior to erythroid commitment Development, June 15, 2008; 135(12): 2071 - 2082. [Abstract] [Full Text] [PDF] A. R. Gordon, S. V. Outram, M. Keramatipour, C. A. Goddard, W. H. Colledge, J. C. Metcalfe, A. L. Hager-Theodorides, T. Crompton, and P. R. Kemp Splenomegaly and Modified Erythropoiesis in KLF13-/- Mice J. Biol. Chem., May 2, 2008; 283(18): 11897 - 11904. [Abstract] [Full Text] [PDF] V. G. Sankaran, S. H. Orkin,, and C. R. Walkley Rb intrinsically promotes erythropoiesis by coupling cell cycle exit with mitochondrial biogenesis Genes & Dev., February 15, 2008; 22(4): 463 - 475. [Abstract] [Full Text] [PDF] S. B. Keel, R. T. Doty, Z. Yang, J. G. Quigley, J. Chen, S. Knoblaugh, P. D. Kingsley, I. De Domenico, M. B. Vaughn, J. Kaplan, et al. A Heme Export Protein Is Required for Red Blood Cell Differentiation and Iron Homeostasis Science, February 8, 2008; 319(5864): 825 - 828. [Abstract] [Full Text] [PDF] G. Yang, S.-C. Huang, J. Y. Wu, and E. J. Benz Jr Regulated Fox-2 isoform expression mediates protein 4.1R splicing during erythroid differentiation Blood, January 1, 2008; 111(1): 392 - 401. [Abstract] [Full Text] [PDF] P. Frontelo, D. Manwani, M. Galdass, H. Karsunky, F. Lohmann, P. G. Gallagher, and J. J. Bieker Novel role for EKLF in megakaryocyte lineage commitment Blood, December 1, 2007; 110(12): 3871 - 3880. [Abstract] [Full Text] [PDF] P. Basu, T. K. Lung, W. Lemsaddek, T. G. Sargent, D. C. Williams Jr, M. Basu, L. C. Redmond, J. B Lingrel, J. L. Haar, and J. A. Lloyd EKLF and KLF2 have compensatory roles in embryonic {beta}-globin gene expression and primitive erythropoiesis Blood, November 1, 2007; 110(9): 3417 - 3425. [Abstract] [Full Text] [PDF] X.-S. Liu, X.-H. Li, Y. Wang, R.-Z. Shu, L. Wang, S.-Y. Lu, H. Kong, Y.-E Jin, L.-J. Zhang, J. Fei, et al. Disruption of palladin leads to defects in definitive erythropoiesis by interfering with erythroblastic island formation in mouse fetal liver Blood, August 1, 2007; 110(3): 870 - 876. [Abstract] [Full Text] [PDF] S. Eshghi, M. G. Vogelezang, R. O. Hynes, L. G. Griffith, and H. F. Lodish {alpha}4{beta}1 integrin and erythropoietin mediate temporally distinct steps in erythropoiesis: integrins in red cell development J. Cell Biol., June 21, 2007; 177(5): 871 - 880. [Abstract] [Full Text] [PDF] J. Zhang, Y. Liu, C. Beard, D. A. Tuveson, R. Jaenisch, T. E. Jacks, and H. F. Lodish Expression of oncogenic K-ras from its endogenous promoter leads to a partial block of erythroid differentiation and hyperactivation of cytokine-dependent signaling pathways Blood, June 15, 2007; 109(12): 5238 - 5241. [Abstract] [Full Text] [PDF] J. Shuga, J. Zhang, L. D. Samson, H. F. Lodish, and L. G. Griffith In vitro erythropoiesis from bone marrow-derived progenitors provides a physiological assay for toxic and mutagenic compounds PNAS, May 22, 2007; 104(21): 8737 - 8742. [Abstract] [Full Text] [PDF] J. M. Perry, O. F. Harandi, and R. F. Paulson BMP4, SCF, and hypoxia cooperatively regulate the expansion of murine stress erythroid progenitors Blood, May 15, 2007; 109(10): 4494 - 4502. [Abstract] [Full Text] [PDF] A. T. Shaw, A. Meissner, J. A. Dowdle, D. Crowley, M. Magendantz, C. Ouyang, T. Parisi, J. Rajagopal, L. J. Blank, R. T. Bronson, et al. Sprouty-2 regulates oncogenic K-ras in lung development and tumorigenesis Genes & Dev., March 15, 2007; 21(6): 694 - 707. [Abstract] [Full Text] [PDF] S. T. Fraser, J. Isern, and M. H. Baron Maturation and enucleation of primitive erythroblasts during mouse embryogenesis is accompanied by changes in cell-surface antigen expression Blood, January 1, 2007; 109(1): 343 - 352. [Abstract] [Full Text] [PDF] D. Yoon, H. Bruchova, A. M. Agarwal, J. T. Prchal, and J. F. Prchal In Vitro Erythroid Cell Expansion Analysis in Polycythemia Vera. Blood (ASH Annual Meeting Abstracts), November 16, 2006; 108(11): 4916 - 4916. [Abstract] [PDF] S. Hosoya-Ohmura, N. Mochizuki, M. Suzuki, O. Ohneda, K. Ohneda, and M. Yamamoto GATA-4 Incompletely Substitutes for GATA-1 in Promoting Both Primitive and Definitive Erythropoiesis in Vivo J. Biol. Chem., October 27, 2006; 281(43): 32820 - 32830. [Abstract] [Full Text] [PDF] B. S. Braun, J. A. Archard, J. A. G. Van Ziffle, D. A. Tuveson, T. E. Jacks, and K. Shannon Somatic activation of a conditional KrasG12D allele causes ineffective erythropoiesis in vivo Blood, September 15, 2006; 108(6): 2041 - 2044. [Abstract] [Full Text] [PDF] B. Dumitriu, M. R. Patrick, J. P. Petschek, S. Cherukuri, U. Klingmuller, P. L. Fox, and V. Lefebvre Sox6 cell-autonomously stimulates erythroid cell survival, proliferation, and terminal maturation and is thereby an important enhancer of definitive erythropoiesis during mouse development Blood, August 15, 2006; 108(4): 1198 - 1207. [Abstract] [Full Text] [PDF] E. Ferretti, J. C. Villaescusa, P. Di Rosa, L. C. Fernandez-Diaz, E. Longobardi, R. Mazzieri, A. Miccio, N. Micali, L. Selleri, G. Ferrari, et al. Hypomorphic Mutation of the TALE Gene Prep1 (pKnox1) Causes a Major Reduction of Pbx and Meis Proteins and a Pleiotropic Embryonic Phenotype Mol. Cell. Biol., August 1, 2006; 26(15): 5650 - 5662. [Abstract] [Full Text] [PDF] Y. Liu, R. Pop, C. Sadegh, C. Brugnara, V. H. Haase, and M. Socolovsky Suppression of Fas-FasL coexpression by erythropoietin mediates erythroblast expansion during the erythropoietic stress response in vivo Blood, July 1, 2006; 108(1): 123 - 133. [Abstract] [Full Text] [PDF] T. Ito, N. Arimitsu, M. Takeuchi, N. Kawamura, M. Nagata, K. Saso, N. Akimitsu, H. Hamamoto, S. Natori, A. Miyajima, et al. Transcription Elongation Factor S-II Is Required for Definitive Hematopoiesis Mol. Cell. Biol., April 15, 2006; 26(8): 3194 - 3203. [Abstract] [Full Text] [PDF] P. G. Gallagher, M. O. Arcasoy, S. E. Vayda, H. K. Dressman, J. J. Bieker, and D. M. Bodine A Differentiation Block in Erythroid Cells Lacking Erythroid Krupple-Like Factor (EKLF). Blood (ASH Annual Meeting Abstracts), November 16, 2005; 106(11): 526 - 526. [Abstract] B. Bonnesen, C. Orskov, S. Rasmussen, P. J. Holst, J. P. Christensen, K. W. Eriksen, K. Qvortrup, N. Odum, and T. Labuda MEK kinase 1 activity is required for definitive erythropoiesis in the mouse fetal liver Blood, November 15, 2005; 106(10): 3396 - 3404. [Abstract] [Full Text] [PDF] J. Zhang and H. F. Lodish Identification of K-ras as the major regulator for cytokine-dependent Akt activation in erythroid progenitors in vivo PNAS, October 11, 2005; 102(41): 14605 - 14610. [Abstract] [Full Text] [PDF] S. Schubbert, K. Lieuw, S. L. Rowe, C. M. Lee, X. Li, M. L. Loh, D. W. Clapp, and K. M. Shannon Functional analysis of leukemia-associated PTPN11 mutations in primary hematopoietic cells Blood, July 1, 2005; 106(1): 311 - 317. [Abstract] [Full Text] [PDF] W. Tong, J. Zhang, and H. F. Lodish Lnk inhibits erythropoiesis and Epo-dependent JAK2 activation and downstream signaling pathways Blood, June 15, 2005; 105(12): 4604 - 4612. [Abstract] [Full Text] [PDF] T. M. Schlaeger, H. K. A. Mikkola, C. Gekas, H. B. Helgadottir, and S. H. Orkin Tie2Cre-mediated gene ablation defines the stem-cell leukemia gene (SCL/tal1)-dependent window during hematopoietic stem-cell development Blood, May 15, 2005; 105(10): 3871 - 3874. [Abstract] [Full Text] [PDF] W. F. Khalaf, H. White, M. J. Wenning, A. Orazi, R. Kapur, and D. A. Ingram K-Ras is essential for normal fetal liver erythropoiesis Blood, May 1, 2005; 105(9): 3538 - 3541. [Abstract] [Full Text] [PDF] G. Terszowski, C. Waskow, P. Conradt, D. Lenze, J. Koenigsmann, D. Carstanjen, I. Horak, and H.-R. Rodewald Prospective isolation and global gene expression analysis of the erythrocyte colony-forming unit (CFU-E) Blood, March 1, 2005; 105(5): 1937 - 1945. [Abstract] [Full Text] [PDF] G. Yang, S.-C. Huang, J. Y. Wu, and E. J. Benz Jr An erythroid differentiation-specific splicing switch in protein 4.1R mediated by the interaction of SF2/ASF with an exonic splicing enhancer Blood, March 1, 2005; 105(5): 2146 - 2153. [Abstract] [Full Text] [PDF] K. W. Harder, C. Quilici, E. Naik, M. Inglese, N. Kountouri, A. Turner, K. Zlatic, D. M. Tarlinton, and M. L. Hibbs Perturbed myelo/erythropoiesis in Lyn-deficient mice is similar to that in mice lacking the inhibitory phosphatases SHP-1 and SHIP-1 Blood, December 15, 2004; 104(13): 3901 - 3910. [Abstract] [Full Text] [PDF] H. Hanawa, P. W. Hargrove, S. Kepes, D. K. Srivastava, A. W. Nienhuis, and D. A. Persons Extended {beta}-globin locus control region elements promote consistent therapeutic expression of a {gamma}-globin lentiviral vector in murine {beta}-thalassemia Blood, October 15, 2004; 104(8): 2281 - 2290. [Abstract] [Full Text] [PDF] T. M. Guadagno Too much ERK, not enough erythrocytes Blood, September 15, 2004; 104(6): 1591 - 1592. [Full Text] [PDF] J. Zhang and H. F. Lodish Constitutive activation of the MEK/ERK pathway mediates all effects of oncogenic H-ras expression in primary erythroid progenitors Blood, September 15, 2004; 104(6): 1679 - 1687. [Abstract] [Full Text] [PDF] J. Antonchuk, C. D. Hyland, D. J. Hilton, and W. S. Alexander Synergistic effects on erythropoiesis, thrombopoiesis, and stem cell competitiveness in mice deficient in thrombopoietin and steel factor receptors Blood, September 1, 2004; 104(5): 1306 - 1313. [Abstract] [Full Text] [PDF] H. Mok, J. Jelinek, S. Pai, B. M. Cattanach, J. T. Prchal, H. Youssoufian, and A. Schumacher Disruption of ferroportin 1 regulation causes dynamic alterations in iron homeostasis and erythropoiesis in polycythaemia mice Development, April 15, 2004; 131(8): 1859 - 1868. [Abstract] [Full Text] [PDF] B. S. Braun, D. A. Tuveson, N. Kong, D. T. Le, S. C. Kogan, J. Rozmus, M. M. Le Beau, T. E. Jacks, and K. M. Shannon Somatic activation of oncogenic Kras in hematopoietic cells initiates a rapidly fatal myeloproliferative disorder PNAS, January 13, 2004; 101(2): 597 - 602. [Abstract] [Full Text] [PDF]

	Copyright © 2003 by American Society of Hematology         Online ISSN: 1528-0020