ETV4 And AP1 Transcription Factors Form Multivalent Interactions With Three Sites On The MED25 Activator-Interacting Domain

Currie, S. L.; Doane, J. J.; Evans, K. S.; Bhachech, N.; Madison, B. J.; Lau, D. K. W.; McIntosh, L. P.; Skalicky, J. J.; Clark, K. A.; Graves, B. J.

doi:10.1101/126458

bioRxiv · 10.1101/126458

ETV4 And AP1 Transcription Factors Form Multivalent Interactions With Three Sites On The MED25 Activator-Interacting Domain

Currie, S. L.·Doane, J. J.·Evans, K. S.·Bhachech, N.·Madison, B. J.·Lau, D. K. W.·McIntosh, L. P.·Skalicky, J. J.·Clark, K. A.·Graves, B. J.

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Abstract

The recruitment of transcriptional cofactors by sequence-specific transcription factors challenges the basis of high affinity and selective interactions. Extending previous studies that the N-terminal activation domain (AD) of ETV5 interacts with Mediator subunit 25 (MED25), we establish that similar, aromatic-rich motifs located both in the AD and in the DNA-binding domain (DBD) of the related ETS factor ETV4 interact with MED25. These ETV4 regions bind MED25 independently, display distinct kinetics, and combine to contribute to a high-affinity interaction of full-length ETV4 with MED25. Within the ETS family, high-affinity interactions with MED25 are specific for the ETV1/4/5 subfamily as other ETS factors display weaker or no detectable binding. The AD binds to a single site on MED25 and the DBD interacts with three MED25 sites, allowing for simultaneous binding of both domains in full-length ETV4. MED25 also stimulates the in vitro DNA binding activity of ETV4 by relieving autoinhibition. ETV1/4/5 factors are often overexpressed in prostate cancer and genome-wide studies in a prostate cancer cell line indicate that ETV4 and MED25 occupy enhancers that are enriched for ETS-binding sequences and are both functionally important for the transcription of genes regulated by these enhancers. AP1-binding sequences were observed in MED25-occupied regions and JUN/FOS also contact MED25; FOS strongly binds to the same MED25 site as ETV4 AD and JUN interacts with the other two MED25 sites. In summary, we describe features of the multivalent ETV4- and AP1-MED25 interactions, thereby implicating these factors in the recruitment of MED25 to transcriptional control elements.

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Currie, S. L., Doane, J. J., Evans, K. S., Bhachech, N., Madison, B. J., Lau, D. K. W., McIntosh, L. P., Skalicky, J. J., Clark, K. A., Graves, B. J.. 2017-04-11. ETV4 And AP1 Transcription Factors Form Multivalent Interactions With Three Sites On The MED25 Activator-Interacting Domain. https://doi.org/10.1101/126458

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