Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Logo for

Science 309 (5731): 142-145

Copyright © 2005 by the American Association for the Advancement of Science

Variable Control of Ets-1 DNA Binding by Multiple Phosphates in an Unstructured Region

Miles A. Pufall,1 Gregory M. Lee,2 Mary L. Nelson,1 Hyun-Seo Kang,2 Algirdas Velyvis,3 Lewis E. Kay,3 Lawrence P. McIntosh,2 Barbara J. Graves1*

Abstract: Cell signaling that culminates in posttranslational modifications directs protein activity. Here we report how multiple Ca2+-dependent phosphorylation sites within the transcription activator Ets-1 act additively to produce graded DNA binding affinity. Nuclear magnetic resonance spectroscopic analyses show that phosphorylation shifts Ets-1 from a dynamic conformation poised to bind DNA to a well-folded inhibited state. These phosphates lie in an unstructured flexible region that functions as the allosteric effector of autoinhibition. Variable phosphorylation thus serves as a "rheostat" for cell signaling to fine-tune transcription at the level of DNA binding.

1 Huntsman Cancer Institute, Department of Oncological Sciences, University of Utah, Salt Lake City, UT 84112–5550, USA.
2 Department of Biochemistry and Molecular Biology, Department of Chemistry, and The Michael Smith Laboratory, University of British Columbia, Vancouver, British Columbia, V6T 1Z3, Canada.
3 Departments of Medical Genetics, Biochemistry, and Chemistry, University of Toronto, Toronto, Ontario, M5S 1A8, Canada.

* To whom correspondence should be addressed. E-mail: Barbara.Graves{at}

Comparative analysis of Erk phosphorylation suggests a mixed strategy for measuring phospho-form distributions.
S. Prabakaran, R. A. Everley, I. Landrieu, J.-M. Wieruszeski, G. Lippens, H. Steen, and J. Gunawardena (2014)
Mol Syst Biol 7, 482
   Abstract »    Full Text »    PDF »
Calcineurin Regulates Nuclear Factor I Dephosphorylation and Activity in Malignant Glioma Cell Lines.
M. Brun, D. D. Glubrecht, S. Baksh, and R. Godbout (2013)
J. Biol. Chem. 288, 24104-24115
   Abstract »    Full Text »    PDF »
CK2 Phosphorylation Inactivates DNA Binding by the Papillomavirus E1 and E2 Proteins.
S. Schuck, C. Ruse, and A. Stenlund (2013)
J. Virol. 87, 7668-7679
   Abstract »    Full Text »    PDF »
Tyrosyl phosphorylation toggles a Runx1 switch.
B. G. Neel and N. A. Speck (2012)
Genes & Dev. 26, 1520-1526
   Abstract »    Full Text »    PDF »
Confirming the Functional Importance of a Protein-DNA Interaction.
M. F. Carey, C. L. Peterson, and S. T. Smale (2012)
Cold Spring Harb Protoc 2012, pdb.top070060
   Abstract »    Full Text »    PDF »
Structural activation of the transcriptional repressor EthR from Mycobacterium tuberculosis by single amino acid change mimicking natural and synthetic ligands.
X. Carette, N. Blondiaux, E. Willery, S. Hoos, N. Lecat-Guillet, Z. Lens, A. Wohlkonig, R. Wintjens, S. H. Soror, F. Frenois, et al. (2012)
Nucleic Acids Res. 40, 3018-3030
   Abstract »    Full Text »    PDF »
ETS1 transcriptional activity is increased in advanced prostate cancer and promotes the castrate-resistant phenotype.
A. M. Smith, V. J. Findlay, S. G. Bandurraga, E. Kistner-Griffin, L. S. Spruill, A. Liu, A. R. Golshayan, and D. P. Turner (2012)
Carcinogenesis 33, 572-580
   Abstract »    Full Text »    PDF »
Identification of cis elements necessary for glucocorticoid induction of growth hormone gene expression in chicken embryonic pituitary cells.
K. Heuck-Knubel, M. Proszkowiec-Weglarz, J. Narayana, L. E. Ellestad, N. Prakobsaeng, and T. E. Porter (2012)
Am J Physiol Regulatory Integrative Comp Physiol 302, R606-R619
   Abstract »    Full Text »    PDF »
Interaction of Ets-1 with HDAC1 Represses IL-10 Expression in Th1 Cells.
C.-G. Lee, H.-K. Kwon, A. Sahoo, W. Hwang, J.-S. So, J.-S. Hwang, C.-S. Chae, G.-C. Kim, J.-E. Kim, H.-S. So, et al. (2012)
J. Immunol. 188, 2244-2253
   Abstract »    Full Text »    PDF »
Phosphorylation of a PDZ Domain Extension Modulates Binding Affinity and Interdomain Interactions in Postsynaptic Density-95 (PSD-95) Protein, a Membrane-associated Guanylate Kinase (MAGUK).
J. Zhang, C. M. Petit, D. S. King, and A. L. Lee (2011)
J. Biol. Chem. 286, 41776-41785
   Abstract »    Full Text »    PDF »
Cell cycle-regulated multi-site phosphorylation of Neurogenin 2 coordinates cell cycling with differentiation during neurogenesis.
F. Ali, C. Hindley, G. McDowell, R. Deibler, A. Jones, M. Kirschner, F. Guillemot, and A. Philpott (2011)
Development 138, 4267-4277
   Abstract »    Full Text »    PDF »
c-Abl-Mediated Tyrosine Phosphorylation of the T-bet DNA-Binding Domain Regulates CD4+ T-Cell Differentiation and Allergic Lung Inflammation.
A. Chen, S.-M. Lee, B. Gao, S. Shannon, Z. Zhu, and D. Fang (2011)
Mol. Cell. Biol. 31, 3445-3456
   Abstract »    Full Text »    PDF »
Tunable Membrane Binding of the Intrinsically Disordered Dehydrin Lti30, a Cold-Induced Plant Stress Protein.
S. K. Eriksson, M. Kutzer, J. Procek, G. Grobner, and P. Harryson (2011)
PLANT CELL 23, 2391-2404
   Abstract »    Full Text »    PDF »
Dynamically committed, uncommitted, and quenched states encoded in protein kinase A revealed by NMR spectroscopy.
L. R. Masterson, L. Shi, E. Metcalfe, J. Gao, S. S. Taylor, and G. Veglia (2011)
PNAS 108, 6969-6974
   Abstract »    Full Text »    PDF »
Graded enhancement of p53 binding to CREB-binding protein (CBP) by multisite phosphorylation.
C. W. Lee, J. C. Ferreon, A. C. M. Ferreon, M. Arai, and P. E. Wright (2010)
PNAS 107, 19290-19295
   Abstract »    Full Text »    PDF »
Control of Activating Transcription Factor 4 (ATF4) Persistence by Multisite Phosphorylation Impacts Cell Cycle Progression and Neurogenesis.
C. L. Frank, X. Ge, Z. Xie, Y. Zhou, and L.-H. Tsai (2010)
J. Biol. Chem. 285, 33324-33337
   Abstract »    Full Text »    PDF »
Local control of a disorder-order transition in 4E-BP1 underpins regulation of translation via eIF4E.
S. Tait, K. Dutta, D. Cowburn, J. Warwicker, A. J. Doig, and J. E. G. McCarthy (2010)
PNAS 107, 17627-17632
   Abstract »    Full Text »    PDF »
Transcriptional silencing of the ETS1 oncogene contributes to human granulocytic differentiation.
V. Lulli, P. Romania, R. Riccioni, A. Boe, F. Lo-Coco, U. Testa, and G. Marziali (2010)
Haematologica 95, 1633-1641
   Abstract »    Full Text »    PDF »
Genome-wide analysis of ETS-family DNA-binding in vitro and in vivo.
G.-H. Wei, G. Badis, M. F. Berger, T. Kivioja, K. Palin, M. Enge, M. Bonke, A. Jolma, M. Varjosalo, A. R. Gehrke, et al. (2010)
EMBO J. 29, 2147-2160
   Abstract »    Full Text »    PDF »
DNA Binding by the ETS Protein TEL (ETV6) Is Regulated by Autoinhibition and Self-association.
S. M. Green, H. J. Coyne III, L. P. McIntosh, and B. J. Graves (2010)
J. Biol. Chem. 285, 18496-18504
   Abstract »    Full Text »    PDF »
Emergent Properties of EWS/FLI Regulation via GGAA Microsatellites in Ewing's Sarcoma.
K. Gangwal, D. Close, C. A. Enriquez, C. P. Hill, and S. L. Lessnick (2010)
Genes & Cancer 1, 177-187
   Abstract »    Full Text »    PDF »
NMR evidence for differential phosphorylation-dependent interactions in WT and {Delta}F508 CFTR.
V. Kanelis, R. P. Hudson, P. H. Thibodeau, P. J. Thomas, and J. D. Forman-Kay (2010)
EMBO J. 29, 263-277
   Abstract »    Full Text »    PDF »
Regulating transcription regulators via allostery and flexibility.
D. Beckett (2009)
PNAS 106, 22035-22036
   Full Text »    PDF »
The transcription factor Ets1 is important for CD4 repression and Runx3 up-regulation during CD8 T cell differentiation in the thymus.
M. Zamisch, L. Tian, R. Grenningloh, Y. Xiong, K. F. Wildt, M. Ehlers, I-C. Ho, and R. Bosselut (2009)
J. Exp. Med. 206, 2685-2699
   Abstract »    Full Text »    PDF »
An age-related homeostasis mechanism is essential for spontaneous amelioration of hemophilia B Leyden.
S. Kurachi, J. S. Huo, A. Ameri, K. Zhang, A. C. Yoshizawa, and K. Kurachi (2009)
PNAS 106, 7921-7926
   Abstract »    Full Text »    PDF »
Regulation of C-type Lectin Antimicrobial Activity by a Flexible N-terminal Prosegment.
S. Mukherjee, C. L. Partch, R. E. Lehotzky, C. V. Whitham, H. Chu, C. L. Bevins, K. H. Gardner, and L. V. Hooper (2009)
J. Biol. Chem. 284, 4881-4888
   Abstract »    Full Text »    PDF »
A coordinated phosphorylation cascade initiated by p38MAPK/MSK1 directs RAR{alpha} to target promoters.
N. Bruck, D. Vitoux, C. Ferry, V. Duong, A. Bauer, H. de The, and C. Rochette-Egly (2009)
EMBO J. 28, 34-47
   Abstract »    Full Text »    PDF »
Dynamic equilibrium engagement of a polyvalent ligand with a single-site receptor.
T. Mittag, S. Orlicky, W.-Y. Choy, X. Tang, H. Lin, F. Sicheri, L. E. Kay, M. Tyers, and J. D. Forman-Kay (2008)
PNAS 105, 17772-17777
   Abstract »    Full Text »    PDF »
Regulation of the transcription factor Ets-1 by DNA-mediated homo-dimerization.
E. P. Lamber, L. Vanhille, L. C. Textor, G. S. Kachalova, M. H. Sieweke, and M. Wilmanns (2008)
EMBO J. 27, 2006-2017
   Abstract »    Full Text »    PDF »
The RCI server: rapid and accurate calculation of protein flexibility using chemical shifts.
M. V. Berjanskii and D. S. Wishart (2007)
Nucleic Acids Res. 35, W531-W537
   Abstract »    Full Text »    PDF »
Polyelectrostatic interactions of disordered ligands suggest a physical basis for ultrasensitivity.
M. Borg, T. Mittag, T. Pawson, M. Tyers, J. D. Forman-Kay, and H. S. Chan (2007)
PNAS 104, 9650-9655
   Abstract »    Full Text »    PDF »
Bidirectional Activity-Dependent Regulation of Neuronal Ion Channel Phosphorylation.
H. Misonou, M. Menegola, D. P. Mohapatra, L. K. Guy, K.-S. Park, and J. S. Trimmer (2006)
J. Neurosci. 26, 13505-13514
   Abstract »    Full Text »    PDF »
Fli1 and Ets1 Have Distinct Roles in Connective Tissue Growth Factor/CCN2 Gene Regulation and Induction of the Profibrotic Gene Program.
S. S. Nakerakanti, B. Kapanadze, M. Yamasaki, M. Markiewicz, and M. Trojanowska (2006)
J. Biol. Chem. 281, 25259-25269
   Abstract »    Full Text »    PDF »
Graded regulation of the Kv2.1 potassium channel by variable phosphorylation..
K.-S. Park, D. P. Mohapatra, H. Misonou, and J. S. Trimmer (2006)
Science 313, 976-979
   Abstract »    Full Text »    PDF »
The Stimulus-induced Tyrosine Phosphorylation of Munc18c Facilitates Vesicle Exocytosis.
E. Oh and D. C. Thurmond (2006)
J. Biol. Chem. 281, 17624-17634
   Abstract »    Full Text »    PDF »
Phosphorylation by PKA potentiates retinoic acid receptor {alpha} activity by means of increasing interaction with and phosphorylation by cyclin H/cdk7.
E. Gaillard, N. Bruck, Y. Brelivet, G. Bour, S. Lalevee, A. Bauer, O. Poch, D. Moras, and C. Rochette-Egly (2006)
PNAS 103, 9548-9553
   Abstract »    Full Text »    PDF »
Cooperative DNA Binding with AP-1 Proteins Is Required for Transformation by EWS-Ets Fusion Proteins.
S. Kim, C. T. Denny, and R. Wisdom (2006)
Mol. Cell. Biol. 26, 2467-2478
   Abstract »    Full Text »    PDF »
Transcription factor ETS1 is critical for human uterine decidualization.
C. A. Kessler, J. K. Schroeder, A. K. Brar, and S. Handwerger (2006)
Mol. Hum. Reprod. 12, 71-76
   Abstract »    Full Text »    PDF »
Can You Hear Me Now? Regulating Transcriptional Activators by Phosphorylation.
K. H. Gardner and M. Montminy (2005)
Sci. STKE 2005, pe44
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882