Abstract
Post-translational modifications provide sensitive and flexible mechanisms to dynamically modulate protein function in response to specific signalling inputs1. In the case of transcription factors, changes in phosphorylation state can influence protein stability, conformation, subcellular localization, cofactor interactions, transactivation potential and transcriptional output1. Here we show that the evolutionarily conserved transcription factor Eyes absent (Eya)2,3 belongs to the phosphatase subgroup of the haloacid dehalogenase (HAD) superfamily4,5, and propose a function for it as a non-thiol-based protein tyrosine phosphatase. Experiments performed in cultured Drosophila cells and in vitro indicate that Eyes absent has intrinsic protein tyrosine phosphatase activity and can autocatalytically dephosphorylate itself. Confirming the biological significance of this function, mutations that disrupt the phosphatase active site severely compromise the ability of Eyes absent to promote eye specification and development in Drosophila. Given the functional importance of phosphorylation-dependent modulation of transcription factor activity, this evidence for a nuclear transcriptional coactivator with intrinsic phosphatase activity suggests an unanticipated method of fine-tuning transcriptional regulation.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Hunter, T. Signaling—2000 and beyond. Cell 100, 113–127 (2000)
Treisman, J. E. A conserved blueprint for the eye? Bioessays 21, 843–850 (1999)
Wawersik, S & Maas, R. L. Vertebrate eye development as modeled in Drosophila. Hum. Mol. Genet. 9, 917–925 (2000)
Collet, J. F., van Schaftingen, E. & Stroobant, V. A new family of phosphotransferases related to P-type ATPases. Trends Biochem. Sci. 23, 284 (1998)
Thaller, M. C., Schippa, S. & Rossolini, G. M. Conserved sequence motifs among bacterial, eukaryotic, and archaeal phosphatases that define a new phosphohydrolase superfamily. Protein Sci. 7, 1647–1652 (1998)
Bonini, N. M., Bui, Q. T., Gray-Board, G. L. & Warrick, J. M. The Drosophila eyes absent gene directs ectopic eye formation in a pathway conserved between flies and vertebrates. Development 124, 4819–4826 (1997)
Pignoni, F. et al. The eye-specification proteins So and Eya form a complex and regulate multiple steps in Drosophila eye development. Cell 91, 881–891 (1997)
Chen, R., Amoui, M., Zhang, Z. & Mardon, G. Dachshund and Eyes absent proteins form a complex and function synergistically to induce ectopic eye development in Drosophila. Cell 91, 893–903 (1997)
Silver, S. J., Davies, E. L., Doyon, L. & Rebay, I. A functional dissection of Eyes absent reveals new modes of regulation within the retinal determination gene network. Mol. Cell. Biol. 23, 5989–5999 (2003)
Kim, S. S. et al. Structure of the retinal determination protein Dachshund reveals a DNA binding motif. Structure (Camb.) 10, 787–795 (2002)
Ikeda, K., Watanabe, Y., Ohto, H. & Kawakami, K. Molecular interaction and synergistic activation of a promoter by Six, Eya, and Dach proteins mediated through CREB binding protein. Mol. Cell. Biol. 22, 6759–6766 (2002)
Bui, Q. T., Zimmerman, J. E., Liu, H. & Bonini, N. M. Molecular analysis of Drosophila eyes absent mutants reveals features of the conserved Eya domain. Genetics 155, 709–720 (2000)
Collet, J. F., Stroobant, V. & Van Schaftingen, E. Mechanistic studies of phosphoserine phosphatase, an enzyme related to P-type ATPases. J. Biol. Chem. 274, 33985–33990 (1999)
Aravind, L., Galperin, M. Y. & Koonin, E. V. The catalytic domain of the P-type ATPase has the haloacid dehalogenase fold. Trends Biochem. Sci. 23, 127–129 (1998)
Selengut, J. D. Mdp-1 is a new and distinct member of the haloacid dehalogenase family of aspartate-dependent phosphohydrolases. Biochemistry 40, 12704–12711 (2001)
Ridder, I. & Dijkstra, B. Identification of the Mg2+ -binding site in the P-type ATPase and phosphatase members of the HAD (haloacid dehalogenase) superfamily by structural similarity to the response regulator protein CheY. Biochem. J. 339, 223–226 (1999)
Cho, H. et al. Beryllium fluoride acts as a phosphate analog in proteins phosphorylated on aspartate: structure of a beryllium fluoride complex with phosphoserine phosphatase. Proc. Natl Acad. Sci. USA 98, 8525–8530 (2001)
Selengut, J. D. & Levine, R. L. MDP-1: A novel eukaryotic magnesium-dependent phosphatase. Biochemistry 39, 8315–8324 (2000)
Andersen, J. N. et al. Structural and evolutionary relationships among protein tyrosine phosphatase domains. Mol. Cell. Biol. 21, 7117–7136 (2001)
Hsiao, F. C., Williams, A., Davies, E. L. & Rebay, I. Eyes absent mediates cross-talk between retinal determination genes and the receptor tyrosine kinase signaling pathway. Dev. Cell 1, 51–61 (2001)
Tootle, T. L., Lee, P. S. & Rebay, I. CRM1-mediated nuclear export and regulated activity of the receptor tyrosine kinase antagonist YAN require specific interactions with MAE. Development 130, 845–857 (2003)
Cohen, J., Altaratz, H., Zick, Y., Klingmuller, U. & Neumann, D. Phosphorylation of erythropoietin receptors in the endoplasmic reticulum by pervanadate-mediated inhibition of tyrosine phosphatases. Biochem. J. 327, 391–397 (1997)
Huyer, G. et al. Mechanism of inhibition of protein-tyrosine phosphatases by vanadate and pervanadate. J. Biol. Chem. 272, 843–851 (1997)
Imbert, V. et al. Induction of tyrosine phosphorylation and T-cell activation by vanadate peroxide, an inhibitor of protein tyrosine phosphatases. Biochem. J. 297, 163–173 (1994)
Ruff, S. J., Chen, K. & Cohen, S. Peroxovanadate induces tyrosine phosphorylation of multiple signaling proteins in mouse liver and kidney. J. Biol. Chem. 272, 1263–1267 (1997)
Scanga, S. E. et al. The conserved PI3K/PTEN/Akt signaling pathway regulates both cell size and survival in Drosophila. Oncogene 19, 3971–3977 (2000)
O'Neill, E. M., Rebay, I., Tjian, R. & Rubin, G. M. The activities of two Ets-related transcription factors required for Drosophila eye development are modulated by the Ras/MAPK pathway. Cell 78, 137–147 (1994)
Aravind, L. & Koonin, E. V. The HD domain defines a new superfamily of metal-dependent phosphohydrolases. Trends Biochem. Sci. 23, 469–472 (1998)
Acknowledgements
We thank M. Voas, D. Doroquez, L. Doyon and B. Chaffee for technical assistance, J. Flynn, S. Neher and S. Flaugh for advice, reagents and technical help, D. Maas for murine Eya complementary DNAs, the Rebay lab for advice and discussions, F. Pignoni for the eya2 stocks used in the rescue assay, and R. Hegde for sharing unpublished information. We are grateful for advice, comments and encouragement from R. Fehon, G. Fink, T. Orr-Weaver, S. Shenolikar, F. Solomon and J. York. T.L.T., S.J.S., I.A.M. and B.E.W.P. are supported by the Ludwig Foundation, the Howard Hughes Medical Institute, the Packard Foundation and the National Science Foundation, respectively. I.R. is supported by the National Eye Institute and the Burroughs Wellcome Fund.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare that they have no competing financial interests.
Supplementary information
Rights and permissions
About this article
Cite this article
Tootle, T., Silver, S., Davies, E. et al. The transcription factor Eyes absent is a protein tyrosine phosphatase. Nature 426, 299–302 (2003). https://doi.org/10.1038/nature02097
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nature02097
This article is cited by
-
EYA4 promotes breast cancer progression and metastasis through its role in replication stress avoidance
Molecular Cancer (2023)
-
The EYA3 tyrosine phosphatase activity promotes pulmonary vascular remodeling in pulmonary arterial hypertension
Nature Communications (2019)
-
EYA4 inhibits hepatocellular carcinoma growth and invasion by suppressing NF‐κB‐dependent RAP1 transactivation
Cancer Communications (2018)
-
ZNF506-dependent positive feedback loop regulates H2AX signaling after DNA damage
Nature Communications (2018)
-
WDR1 is a novel EYA3 substrate and its dephosphorylation induces modifications of the cellular actin cytoskeleton
Scientific Reports (2018)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.