The long-term objectives of the proposed research are to understand how specificity is determined within a family of eukaryotic regulatory transcription factors. The ets gene family, which is found throughout the metazoa, has 27 human paralogs. ETS proteins have highly conserved DNA binding properties, yet display unique distinct biological properties. Thus, specificity must have evolved to direct these proteins to function in unique biological contexts. This proposal focuses on how signaling pathways affect the function of ETS proteins to give distinct activities.
Aim I investigates the autoinhibition of Ets-1 DNA binding, including reinforcement by calciumdependent phosphorylation. Biochemical and biophysical approaches will test the role of protein dynamics and conformational change in the regulation of Ets-1 DNA binding. A phosphorylation-dependent rheostat model of Ets-1 regulation will be tested in vitro and in cells.
Aim II investigates Ets-1 and highly-related Ets-2 as effectors of Ras/MAPKinase signaling. We will focus on the macromolecular interfaces that mediate the docking of the MAPK ERK2 to Ets-1 and Ets-2 and the phosphorylation-dependent recruitment of the co-activators CBP/p300. In addition, two recently discovered Ets-1 modifications, acetylation and sumolation, will be investigated.
Aim III will identify authentic transcriptional targets for Ets-1 and Ets-2 to investigate how signaling impinges on the activity of Ets-1 and Ets-2 at genomic loci. We will test the role of post-translational modifications and factor assembly in gene activation and repression. Ras/MAPK signaling and the ETS family of transcription factors regulate genes necessary for control of the cell cycle, apoptosis, and normal differentiation. Mutation of human ras and ets genes contribute to cellular dysfunction in cancer. Indeed, the ras gene is mutated in 20% of human cancers. Therefore, deciphering the molecular function of Ets-1 and Ets-2 within Ras-dependent signal transduction will provide new insight and guide development of new tools to confront the cancer problem.
|Madison, Bethany J; Clark, Kathleen A; Bhachech, Niraja et al. (2018) Electrostatic repulsion causes anticooperative DNA binding between tumor suppressor ETS transcription factors and JUN-FOS at composite DNA sites. J Biol Chem 293:18624-18635|
|Currie, Simon L; Doane, Jedediah J; Evans, Kathryn S et al. (2017) ETV4 and AP1 Transcription Factors Form Multivalent Interactions with three Sites on the MED25 Activator-Interacting Domain. J Mol Biol 429:2975-2995|
|Currie, Simon L; Lau, Desmond K W; Doane, Jedediah J et al. (2017) Structured and disordered regions cooperatively mediate DNA-binding autoinhibition of ETS factors ETV1, ETV4 and ETV5. Nucleic Acids Res 45:2223-2241|
|Desjardins, Geneviève; Okon, Mark; Graves, Barbara J et al. (2016) Conformational Dynamics and the Binding of Specific and Nonspecific DNA by the Autoinhibited Transcription Factor Ets-1. Biochemistry 55:4105-18|
|Kikani, Chintan K; Wu, Xiaoying; Paul, Litty et al. (2016) Pask integrates hormonal signaling with histone modification via Wdr5 phosphorylation to drive myogenesis. Elife 5:|
|De, Soumya; Okon, Mark; Graves, Barbara J et al. (2016) Autoinhibition of ETV6 DNA Binding Is Established by the Stability of Its Inhibitory Helix. J Mol Biol 428:1515-30|
|Huang, Fu; Ramakrishnan, Saravanan; Pokhrel, Srijana et al. (2015) Interaction of the Jhd2 Histone H3 Lys-4 Demethylase with Chromatin Is Controlled by Histone H2A Surfaces and Restricted by H2B Ubiquitination. J Biol Chem 290:28760-77|
|De, Soumya; Chan, Anson C K; Coyne 3rd, H Jerome et al. (2014) Steric mechanism of auto-inhibitory regulation of specific and non-specific DNA binding by the ETS transcriptional repressor ETV6. J Mol Biol 426:1390-406|
|Clark, Kathleen A; Graves, Barbara J (2014) Dual views of SRF: a genomic exposure. Genes Dev 28:926-8|
|Desjardins, Geneviève; Meeker, Charles A; Bhachech, Niraja et al. (2014) Synergy of aromatic residues and phosphoserines within the intrinsically disordered DNA-binding inhibitory elements of the Ets-1 transcription factor. Proc Natl Acad Sci U S A 111:11019-24|
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