Background/Aims: Despite a sophisticated understanding of the hematopoietic developmental program at the transcriptional level, our understanding of the role of E3 ubiquitin ligases remains underdeveloped. The E3 ubiquitin ligase, GRAIL (RNF128), is expressed in the bone marrow, but its role is as yet undefined. In this study, we evaluate the effect of GRAIL expression during hematopoietic differentiation in vitro and in vivo. Methods: Retroviral transduction of hematopoietic multipotent progenitor cells was used for methylcellulose colony assays and bone marrow reconstitution. Results: Enforced expression of GRAIL in colony assays demonstrated skewing of hematopoietic lineage development toward granulocytic/monocytic cells. Bone marrow reconstitution experiments with progenitor cells expressing biologically varied levels of GRAIL demonstrated diminished erythropoiesis and megakaryopoiesis if GRAIL (endogenous or forced) is maintained throughout hematopoiesis. Conclusion: These data highlight a role for GRAIL during hematopoiesis and emphasize the importance of studying posttranslational effects to complement our current understanding of the transcriptional regulation of hematopoiesis.

Keywords:
Hematopoiesis, Hematopoietic progenitor cells, Megakaryocytes thrombocytopenia, Stem cell biology
1.
Cantor AB, Orkin SH: Hematopoietic development: a balancing act. Curr Opin Genet Dev 2001;11:513–519.
2.
Joazeiro CA, Weissman AM: Ring finger proteins: mediators of ubiquitin ligase activity. Cell 2000;102:549–552.
3.
Soares L, Seroogy C, Skrenta H, Anandasabapathy N, Lovelace P, Chung CD, Engleman E, Fathman CG: Two isoforms of otubain 1 regulate T cell anergy via grail. Nat Immunol 2004;5:45–54.
4.
Seroogy CM, Soares L, Ranheim EA, Su L, Holness C, Bloom D, Fathman CG: The gene related to anergy in lymphocytes, an E3 ubiquitin ligase, is necessary for anergy induction in CD4 t cells. J Immunol 2004;173:79–85.
5.
Guais A, Siegrist S, Solhonne B, Jouault H, Guellaen G, Bulle F: H-goliath, paralog of GRAIL, is a new E3 ligase protein, expressed in human leukocytes. Gene 2006;374:112–120.
6.
Van Parijs L, Refaeli Y, Abbas AK, Baltimore D: Autoimmunity as a consequence of retrovirus-mediated expression of C-FLIP in lymphocytes. Immunity 1999;11:763–770.
7.
Anandasabapathy N, Ford G, Bloom D, Holness C, Seroogy C, Skrenta H, Hollenhurst M, Fathman CG, Soares L: Grail: an E3 ubiquitin ligase that inhibits cytokine gene transcription is expressed in anergic CD4+ T cells. Immunity 2003;18:1–20.
8.
Costa GL, Benson JM, Seroogy CM, Achacoso P, Fathman CG, Nolan GP: Targeting rare populations of murine antigen-specific T lymphocytes by retroviral transduction for potential application in gene therapy for autoimmune disease. J Immunol 2000;164:3581–3590.
9.
Mackenzie DA, Schartner J, Lin J, Timmel A, Jennens-Clough M, Fathman CG, Seroogy CM: GRAIL is upregulated in CD4+ CD25+ T regulatory cells and is sufficient for conversion of T cells to a regulatory phenotype. J Biol Chem 2007;282:9696–9702.
10.
Akashi K, Traver D, Miyamoto T, Weissman IL: A clonogenic common myeloid progenitor that gives rise to all myeloid lineages. Nature 2000;404:193–197.
11.
Slayton WB, Georgelas A, Pierce LJ, Elenitoba-Johnson KS, Perry SS, Marx M, Spangrude GJ: The spleen is a major site of megakaryopoiesis following transplantation of murine hematopoietic stem cells. Blood 2002;100:3975–3982.
12.
Lineberry NB, Su LL, Lin JT, Coffey GP, Seroogy CM, Fathman CG: Cutting edge: the transmembrane E3 ligase GRAIL ubiquitinates the costimulatory molecule CD40 ligand during the induction of T cell anergy. J Immunol 2008;181:1622–1626.
13.
Su L, Lineberry N, Huh Y, Soares L, Fathman CG: A novel E3 ubiquitin ligase substrate screen identifies rho guanine dissociation inhibitor as a substrate of gene related to anergy in lymphocytes. J Immunol 2006;177:7559–7566.
14.
Juris SJ, Shah K, Shokat K, Dixon JE, Vacratsis PO: Identification of otubain 1 as a novel substrate for the yersinia protein kinase using chemical genetics and mass spectrometry. FEBS Lett 2006;580:179–183.
15.
Orkin SH: Diversification of haematopoietic stem cells to specific lineages. Nat Rev Genet 2000;1:57–64.
16.
Kondo M, Scherer DC, Miyamoto T, King AG, Akashi K, Sugamura K, Weissman IL: Cell-fate conversion of lymphoid-committed progenitors by instructive actions of cytokines. Nature 2000;407:383–386.
17.
Iwasaki-Arai J, Iwasaki H, Miyamoto T, Watanabe S, Akashi K: Enforced granulocyte/macrophage colony-stimulating factor signals do not support lymphopoiesis, but instruct lymphoid to myelomonocytic lineage conversion. J Exp Med 2003;197:1311–1322.
18.
Zhang Y, Morrone G, Zhang J, Chen X, Lu X, Ma L, Moore M, Zhou P: CUL-4A stimulates ubiquitylation and degradation of the HOXA9 homeodomain protein. Embo J 2003;22:6057–6067.
19.
Liu LQ, Ilaria R Jr, Kingsley PD, Iwama A, van Etten RA, Palis J, Zhang DE: A novel ubiquitin-specific protease, UBP43, cloned from leukemia fusion protein AML1-eto-expressing mice, functions in hematopoietic cell differentiation. Mol Cell Biol 1999;19:3029–3038.
20.
Chen CY, Pajak L, Tamburlin J, Bofinger D, Koury ST: The effect of proteasome inhibitors on mammalian erythroid terminal differentiation. Exp Hematol 2002;30:634–639.
21.
Marteijn JA, Jansen JH, van der Reijden BA: Ubiquitylation in normal and malignant hematopoiesis: novel therapeutic targets. Leukemia 2006;20:1511–1518.
22.
Richardson PG, Sonneveld P, Schuster MW, Irwin D, Stadtmauer EA, Facon T, Harousseau JL, Ben-Yehuda D, Lonial S, Goldschmidt H, Reece D, San-Miguel JF, Blade J, Boccadoro M, Cavenagh J, Dalton WS, Boral AL, Esseltine DL, Porter JB, Schenkein D, Anderson KC: Bortezomib or high-dose dexamethasone for relapsed multiple myeloma. N Engl J Med 2005;352:2487–2498.
© 2009 S. Karger AG, Basel
2009
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.