The proposal investigates transcription factors encoded by the ETS gene family. These regulatory proteins play central roles in a variety of human cancers, including prostate cancer, Ewings sarcoma and various leukemias. DNA binding transcription factors, like those in the ETS family, orchestrate regulation of gene expression by binding to sequence motifs within promoter and enhancer elements and by recruiting additional multi-protein complexes. This important regulatory process is complicated by the conservation of DNA binding properties among related transcription factors. This proposal addresses how the ETS factors find functional sites in the genome and mediate the assembly of both positive- and negative-acting machinery. We employ a combination of biochemical and genomic approaches including NMR-based structure determination and genome-wide occupancy.
Aim 1 will determine the mechanism of autoinhibition of DNA binding in ETS factors and how this on-board repressive activity is used in biological regulation. We will determine the mechanism of ETV6 autoinhibition and how it plays a role in the repression of transcriptional targets. Based on our findings with ETS1 and ETV6, we will next test for autoinhibition of oncogenic ETS factors, ERG and ETV1, and decipher possible regulatory routes for DNA binding.
Aim 2 focuses on the PNT domain, which represents a divergence point in the ETS family serving to individualize family members. We will investigate the three PNT domain-binding factors for the oncogenic ETS factor ERG that implicates this single factor in both negative and positive transcriptional regulation.
Aim 3 investigates the redundant and specific use of the ETS family within cell-specific networks. ETS factors have a redundant function at the proximal promoters of housekeeping genes, yet can display specific properties at enhancers. We will disrupt ETS networks in normal and diseased cell states to decipher rules for redundant versus specific occupancy. We will test for mechanistic roles of ETS factors in regulating chromatin dynamics and DNA methylation with a focus on redundantly-occupied promoter proximal sites. Our investigation on mechanistic aspects of ETS factors will guide future interventional strategies in the cancer setting. Furthermore, our identification of downstream transcriptional targets of oncogenic ETS factors can help elucidate mechanisms of carcinogenesis.
The genetic information in human genome is accessed by the process of transcription of DNA to RNA. DNA binding proteins, such as the ETS factors, regulate this process. The ETS factors can be mutated or over-expressed in human cancers. Thus, the proposed studies not only will discover basic mechanisms of transcriptional regulation that will impact a wide variety of human diseases, but also will impact development of diagnostic, prevention or treatment strategies for human cancer.
|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|
|Clark, Kathleen A; Graves, Barbara J (2014) Dual views of SRF: a genomic exposure. Genes Dev 28:926-8|
|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|
|Nelson, Mary L; Kang, Hyun-Seo; Lee, Gregory M et al. (2010) Ras signaling requires dynamic properties of Ets1 for phosphorylation-enhanced binding to coactivator CBP. Proc Natl Acad Sci U S A 107:10026-31|
|Thyagarajan, Bargavi; Blaszczak, Adam G; Chandler, Katherine J et al. (2010) ETS-4 is a transcriptional regulator of life span in Caenorhabditis elegans. PLoS Genet 6:e1001125|
|Green, Sean M; Coyne 3rd, H Jerome; McIntosh, Lawrence P et al. (2010) DNA binding by the ETS protein TEL (ETV6) is regulated by autoinhibition and self-association. J Biol Chem 285:18496-504|
|Kang, Hyun-Seo; Nelson, Mary L; Mackereth, Cameron D et al. (2008) Identification and structural characterization of a CBP/p300-binding domain from the ETS family transcription factor GABP alpha. J Mol Biol 377:636-46|
|Gangwal, Kunal; Sankar, Savita; Hollenhorst, Peter C et al. (2008) Microsatellites as EWS/FLI response elements in Ewing's sarcoma. Proc Natl Acad Sci U S A 105:10149-54|
|Hollenhorst, Peter C; Shah, Atul A; Hopkins, Christopher et al. (2007) Genome-wide analyses reveal properties of redundant and specific promoter occupancy within the ETS gene family. Genes Dev 21:1882-94|
|Pufall, Miles A; Lee, Gregory M; Nelson, Mary L et al. (2005) Variable control of Ets-1 DNA binding by multiple phosphates in an unstructured region. Science 309:142-5|
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