Enhancers with cooperative Notch binding sites are more resistant to regulation by the Hairless co-repressor

Yi Kuang, Anna Pyo, Natanel Eafergan, Brittany Cain, Lisa M. Gutzwiller, Ofri Axelrod, Ellen K. Gagliani, Matthew T. Weirauch, Raphael Kopan, Rhett A. Kovall, David Sprinzak, Brian Gebelein*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Notch signaling controls many developmental processes by regulating gene expression. Notch-dependent enhancers recruit activation complexes consisting of the Notch intracellular domain, the Cbf/Su(H)/Lag1 (CSL) transcription factor (TF), and the Mastermind co-factor via two types of DNA sites: monomeric CSL sites and cooperative dimer sites called Su (H) paired sites (SPS). Intriguingly, the CSL TF can also bind co-repressors to negatively regulate transcription via these same sites. Here, we tested how synthetic enhancers with monomeric CSL sites versus dimeric SPSs bind Drosophila Su(H) complexes in vitro and mediate transcriptional outcomes in vivo. Our findings reveal that while the Su(H)/Hairless co-repressor complex similarly binds SPS and CSL sites in an additive manner, the Notch activation complex binds SPSs, but not CSL sites, in a cooperative manner. Moreover, transgenic reporters with SPSs mediate stronger, more consistent transcription and are more resistant to increased Hairless co-repressor expression compared to reporters with the same number of CSL sites. These findings support a model in which SPS containing enhancers preferentially recruit cooperative Notch activation complexes over Hairless repression complexes to ensure consistent target gene activation.

Original languageEnglish
Article numbere1009039
JournalPLoS Genetics
Volume17
Issue number9
DOIs
StatePublished - 24 Sep 2021

Funding

FundersFunder number
BinationalScienceFoundation-1715822andNSF/MCB-BSF-2114950
Cincinnati Children's Hospital
MCB-BSF-2114950, 1715822
National Science Foundation
National Institutes of HealthCA178974, NS099068
National Institute of General Medical SciencesR01GM055479
United States-Israel Binational Science Foundation

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