The mammalian E2F family of transcription factors plays a crucial role in the regulation of cellular proliferation, apoptosis and differentiation. E2F7 and E2F8 are the most recently identified family members and have unusual features that distinguish them from other members in the E2F family, including two distinct DNA-binding domains that bind to DNA in a DP-independent manner. E2F7 and E2F8 have been shown to be transcriptional repressors. However, the mechanism by which E2F7 represses E2F responsive gene expression remains to be elucidated. The results presented here provide the first insight into the E2F7-mediated transcriptional mechanism. E2F7 was shown to contain a CtBP binding motif and form a complex with CtBP in both HeLa and MCF7 cells. An E2F7 deletion mutant lacking the CtBP binding motif was unable to form a complex with CtBP and repress the transcription of E2F target genes in luciferase assays, suggesting that this motif is essential for E2F7-dependent repression. Furthermore, the E2F7-CtBP complex was shown to be stable under different types of damage, such as following etoposide and UV treatment, and under different cell redox states. Interestingly, however, E2F7 was unable to repress the transcriptional activity of E2F target genes following treatment with the CtBP substrate MTOB. Moreover, E2F7 endogenous immunoprecipitations showed that E2F7 forms a complex with the chromatin-modifying enzymes HDAC1, HDAC2 and LSD1 and the co-repressor CoREST. Interestingly, via chromatin immunoprecipitations, E2F7 was shown to recruit these co-repressors to a subset of E2F-responsive promoters where they affect the activity of these promoters in a target gene-specific manner. Furthermore, results presented here suggest that CtBP could play a dual role in E2F7 function, not only being involved in E2F7-mediated repression but also in the repression of E2F7 itself as siRNA mediated CtBP depletion was shown to cause an upregulation of E2F7 protein levels. These results implicate a repertoire of co-repressors in a target gene-specific E2F7 repression mechanism, and as such define a new level of complexity in cell cycle control.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:580944 |
| Date | January 2012 |
| Creators | Kesoglidou, Poli Xenia |
| Contributors | La Thangue, Nicholas |
| Publisher | University of Oxford |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://ora.ox.ac.uk/objects/uuid:5694f75b-0e73-493c-a6bd-da10793aded3 |
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