This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow An erratum has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Eisbacher, M.
Right arrow Articles by Chong, B. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Eisbacher, M.
Right arrow Articles by Chong, B. H.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, May 2003, p. 3427-3441, Vol. 23, No. 10
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.10.3427-3441.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Protein-Protein Interaction between Fli-1 and GATA-1 Mediates Synergistic Expression of Megakaryocyte-Specific Genes through Cooperative DNA Binding

Michael Eisbacher,1 Melissa L. Holmes,1 Anthea Newton,2 Philip J. Hogg,1 Levon M. Khachigian,1 Merlin Crossley,2 and Beng H. Chong1,3*

Centre for Thrombosis and Vascular Research, School of Medical Sciences and Department of Medicine, St. George Clinical School, University of New South Wales, Sydney, New South Wales 2052,1 School of Molecular and Microbial Biosciences, University of Sydney, Sydney, New South Wales 2006, Australia, and,2 National Heart Centre, Singapore 168752, Singapore3

Received 16 January 2003/ Accepted

Friend leukemia integration 1 (Fli-1) is a member of the Ets family of transcriptional activators that has been shown to be an important regulator during megakaryocytic differentiation. We undertook a two-hybrid screen of a K562 cDNA library to identify transcription factors that interacted with Fli-1 and were potential regulators of megakaryocyte development. Here we report the physical interaction of Fli-1 with GATA-1, a well-characterized, zinc finger transcription factor critical for both erythroid and megakaryocytic differentiation. We map the minimal domains required for the interaction and show that the zinc fingers of GATA-1 interact with the Ets domain of Fli-1. GATA-1 has previously been shown to interact with the Ets domain of the Fli-1-related protein PU.1, and the two proteins appear to inhibit each other's activity. In contrast, we demonstrate that GATA-1 and Fli-1 synergistically activate the megakaryocyte-specific promoters GPIX and GPIb{alpha} in transient transfections. Quantitative electrophoretic mobility shift assays using oligonucleotides derived from the GPIX promoter containing Ets and GATA binding motifs reveal that Fli-1 and GATA-1 exhibit cooperative DNA binding in which the binding of GATA-1 to DNA is increased approximately 26-fold in the presence of Fli-1 (from 4.2 to 0.16 nM), providing a mechanism for the observed transcriptional synergy. To test the effect on endogenous genes, we stably overexpressed Fli-1 in K562 cells, a line rich in GATA-1. Overexpression of Fli-1 induced the expression of the endogenous GPIX and GPIb{alpha} genes as measured by Northern blot and fluorescence-activated cell sorter analysis. This work suggests that Fli-1 and GATA-1 work together to activate the expression of genes associated with the terminal differentiation of megakaryocytes.

* Corresponding author. Mailing address: Department of Medicine, Level 2, Clinical Sciences Building, St. George Hospital, Kogarah, NSW, Australia 2217. Phone: 61-2-9350-2011. Fax: 61-2-9350-3998. E-mail: beng.chong{at}unsw.edu.au.

Molecular and Cellular Biology, May 2003, p. 3427-3441, Vol. 23, No. 10
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.10.3427-3441.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Stankiewicz, M. J., Crispino, J. D. (2009). ETS2 and ERG promote megakaryopoiesis and synergize with alterations in GATA-1 to immortalize hematopoietic progenitor cells. Blood 113: 3337-3347 [Abstract] [Full Text]  
  • Matsubara, E., Sakai, I., Yamanouchi, J., Fujiwara, H., Yakushijin, Y., Hato, T., Shigemoto, K., Yasukawa, M. (2009). The Role of Zinc Finger Protein 521/Early Hematopoietic Zinc Finger Protein in Erythroid Cell Differentiation. J. Biol. Chem. 284: 3480-3487 [Abstract] [Full Text]  
  • Bouilloux, F., Juban, G., Cohet, N., Buet, D., Guyot, B., Vainchenker, W., Louache, F., Morle, F. (2008). EKLF restricts megakaryocytic differentiation at the benefit of erythrocytic differentiation. Blood 112: 576-584 [Abstract] [Full Text]  
  • Woo, A. J., Moran, T. B., Schindler, Y. L., Choe, S.-K., Langer, N. B., Sullivan, M. R., Fujiwara, Y., Paw, B. H., Cantor, A. B. (2008). Identification of ZBP-89 as a Novel GATA-1-Associated Transcription Factor Involved in Megakaryocytic and Erythroid Development. Mol. Cell. Biol. 28: 2675-2689 [Abstract] [Full Text]  
  • Viger, R. S., Guittot, S. M., Anttonen, M., Wilson, D. B., Heikinheimo, M. (2008). Role of the GATA Family of Transcription Factors in Endocrine Development, Function, and Disease. Mol. Endocrinol. 22: 781-798 [Abstract] [Full Text]  
  • Fuhrken, P. G., Chen, C., Apostolidis, P. A., Wang, M., Miller, W. M., Papoutsakis, E. T. (2008). Gene Ontology-driven transcriptional analysis of CD34+ cell-initiated megakaryocytic cultures identifies new transcriptional regulators of megakaryopoiesis. Physiol. Genomics 33: 159-169 [Abstract] [Full Text]  
  • Frontelo, P., Manwani, D., Galdass, M., Karsunky, H., Lohmann, F., Gallagher, P. G., Bieker, J. J. (2007). Novel role for EKLF in megakaryocyte lineage commitment. Blood 110: 3871-3880 [Abstract] [Full Text]  
  • Wang, R.-S., Wang, Y., Zhang, X.-S., Chen, L. (2007). Inferring transcriptional regulatory networks from high-throughput data. Bioinformatics 23: 3056-3064 [Abstract] [Full Text]  
  • Nurden, A. T., Nurden, P. (2007). Inherited thrombocytopenias. haematol 92: 1158-1164 [Full Text]  
  • Chan, W. Y. I., Follows, G. A., Lacaud, G., Pimanda, J. E., Landry, J.-R., Kinston, S., Knezevic, K., Piltz, S., Donaldson, I. J., Gambardella, L., Sablitzky, F., Green, A. R., Kouskoff, V., Gottgens, B. (2007). The paralogous hematopoietic regulators Lyl1 and Scl are coregulated by Ets and GATA factors, but Lyl1 cannot rescue the early Scl-/- phenotype. Blood 109: 1908-1916 [Abstract] [Full Text]  
  • Pang, L., Xue, H.-H., Szalai, G., Wang, X., Wang, Y., Watson, D. K., Leonard, W. J., Blobel, G. A., Poncz, M. (2006). Maturation stage-specific regulation of megakaryopoiesis by pointed-domain Ets proteins. Blood 108: 2198-2206 [Abstract] [Full Text]  
  • Liew, C. W., Rand, K. D., Simpson, R. J. Y., Yung, W. W., Mansfield, R. E., Crossley, M., Proetorius-Ibba, M., Nerlov, C., Poulsen, F. M., Mackay, J. P. (2006). Molecular Analysis of the Interaction between the Hematopoietic Master Transcription Factors GATA-1 and PU.1. J. Biol. Chem. 281: 28296-28306 [Abstract] [Full Text]  
  • Maeda, K., Nishiyama, C., Tokura, T., Nakano, H., Kanada, S., Nishiyama, M., Okumura, K., Ogawa, H. (2006). FOG-1 represses GATA-1-dependent Fc{epsilon}RI beta-chain transcription: transcriptional mechanism of mast-cell-specific gene expression in mice. Blood 108: 262-269 [Abstract] [Full Text]  
  • Guyot, B., Murai, K., Fujiwara, Y., Valverde-Garduno, V., Hammett, M., Wells, S., Dear, N., Orkin, S. H., Porcher, C., Vyas, P. (2006). Characterization of a Megakaryocyte-specific Enhancer of the Key Hemopoietic Transcription Factor GATA1. J. Biol. Chem. 281: 13733-13742 [Abstract] [Full Text]  
  • van den Akker, E., Ano, S., Shih, H.-M., Wang, L.-C., Pironin, M., Palvimo, J. J., Kotaja, N., Kirsh, O., Dejean, A., Ghysdael, J. (2005). FLI-1 Functionally Interacts with PIASx{alpha}, a Member of the PIAS E3 SUMO Ligase Family. J. Biol. Chem. 280: 38035-38046 [Abstract] [Full Text]  
  • Bonello, M. R., Bobryshev, Y. V., Khachigian, L. M. (2005). Peroxide-Inducible Ets-1 Mediates Platelet-Derived Growth Factor Receptor-{alpha} Gene Transcription in Vascular Smooth Muscle Cells. Am. J. Pathol. 167: 1149-1159 [Abstract] [Full Text]  
  • Huang, D.-Y., Kuo, Y.-Y., Chang, Z.-F. (2005). GATA-1 mediates auto-regulation of Gfi-1B transcription in K562 cells. Nucleic Acids Res 33: 5331-5342 [Abstract] [Full Text]  
  • Geng, Y., Laslo, P., Barton, K., Wang, C.-R. (2005). Transcriptional Regulation of CD1D1 by Ets Family Transcription Factors. J. Immunol. 175: 1022-1029 [Abstract] [Full Text]  
  • Hughan, S. C., Senis, Y., Best, D., Thomas, A., Frampton, J., Vyas, P., Watson, S. P. (2005). Selective impairment of platelet activation to collagen in the absence of GATA1. Blood 105: 4369-4376 [Abstract] [Full Text]  
  • Hu, W., Philips, A. S., Kwok, J. C., Eisbacher, M., Chong, B. H. (2005). Identification of Nuclear Import and Export Signals within Fli-1: Roles of the Nuclear Import Signals in Fli-1-Dependent Activation of Megakaryocyte-Specific Promoters. Mol. Cell. Biol. 25: 3087-3108 [Abstract] [Full Text]  
  • Masuya, M., Moussa, O., Abe, T., Deguchi, T., Higuchi, T., Ebihara, Y., Spyropoulos, D. D., Watson, D. K., Ogawa, M. (2005). Dysregulation of granulocyte, erythrocyte, and NK cell lineages in Fli-1 gene-targeted mice. Blood 105: 95-102 [Abstract] [Full Text]  
  • Jackers, P., Szalai, G., Moussa, O., Watson, D. K. (2004). Ets-dependent Regulation of Target Gene Expression during Megakaryopoiesis. J. Biol. Chem. 279: 52183-52190 [Abstract] [Full Text]  
  • Elagib, K. E., Xiao, M., Hussaini, I. M., Delehanty, L. L., Palmer, L. A., Racke, F. K., Birrer, M. J., Shanmugasundaram, G., McDevitt, M. A., Goldfarb, A. N. (2004). Jun Blockade of Erythropoiesis: Role for Repression of GATA-1 by HERP2. Mol. Cell. Biol. 24: 7779-7794 [Abstract] [Full Text]  
  • Shimizu, R., Ohneda, K., Engel, J. D., Trainor, C. D., Yamamoto, M. (2004). Transgenic rescue of GATA-1-deficient mice with GATA-1 lacking a FOG-1 association site phenocopies patients with X-linked thrombocytopenia. Blood 103: 2560-2567 [Abstract] [Full Text]  
  • Ghirlando, R., Trainor, C. D. (2003). Determinants of GATA-1 Binding to DNA: THE ROLE OF NON-FINGER RESIDUES. J. Biol. Chem. 278: 45620-45628 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»