BEN factors are conserved CSL co-repressors in Notch-mediated neural development. Our general goal is to understand how cell signaling pathways mediate accurate transcriptional gene control during development. We focus on the Notch signaling pathway and its roles in directing neural cell fates and states. We strive to elucidate general principles of gene regulation by combining studies in both Drosophila and mammalian model systems. This proposal describes first in vivo functional studies of BEN-solo domain proteins. From our Drosophila work, we found that the BEN-solo factor Insensitive (Insv). regulates Notch signaling and peripheral nervous system development by acting as a direct corepressor for the Notch transcription factor CSL. We will further study its genetic requirements, dissect its functional domains, and elucidate how it cooperates with other chromatin factors to induce transcriptional repression. We build upon this proprietary knowledge to study roles of mammalian BEN-solo factors during neural development, and we have preliminary data on their ability to inhibit Notch signaling and affect neural stem cell self-renewal and differentiation. These phenotype-driven studies will be complemented by genomewide analyses of chromatin binding of CSL and BEN-solo factors in flies and mice, from which we will extract Notch-regulated enhancers. We will validate enhancers in vivo with emphasis on neural regulatory elements. We hope to determine conserved features of N/CSL/BEN target networks in the nervous system. In addition, these data potentially shed light on CSL-independent association of BEN-solo proteins with chromatin, which we will address using functional assays. Overall, this proposal combines a variety of experimental approaches and model systems to investigate a novel conserved neural corepressor in the Notch pathway.

Public Health Relevance

Normal development requires cell-cell signaling and precise control over gene regulation. We study this with respect to the Notch pathway, a cell communication system that is fundamental to establishment of cell fates, tissue patterning, and gene control. Through our studies of fruitfly mechanosensory bristles, which require Notch signaling for their accurate development, we found that a BEN family protein regulates Notch signaling and bristle development. This factor associates with chromatin and binds the Notch transcription factor to oppose Notch signaling. Learning from these studies, we engaged first studies of related BEN proteins in the mammalian nervous system, and how they affect the self-renewal of neural stem cells in the embryo and adult. Finally, we are creating genomewide maps of the locations of the Notch transcription factor and BEN proteins. These data yield genetic, biochemical, and bioinformatic insights into the function of a novel conserved family of Notch regulators, and their roles in gene regulatory networks underlying neural development.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS074037-02
Application #
8325028
Study Section
Neurogenesis and Cell Fate Study Section (NCF)
Program Officer
Riddle, Robert D
Project Start
2011-09-01
Project End
2016-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
2
Fiscal Year
2012
Total Cost
$427,562
Indirect Cost
$193,794
Name
Sloan-Kettering Institute for Cancer Research
Department
Type
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10065
Miura, Pedro; Sanfilippo, Piero; Shenker, Sol et al. (2014) Alternative polyadenylation in the nervous system: to what lengths will 3' UTR extensions take us? Bioessays 36:766-77
Aoki, Tsutomu; Wolle, Daniel; Preger-Ben Noon, Ella et al. (2014) Bi-functional cross-linking reagents efficiently capture protein-DNA complexes in Drosophila embryos. Fly (Austin) 8:43-51
Milton, Claire C; Grusche, Felix A; Degoutin, Joffrey L et al. (2014) The Hippo pathway regulates hematopoiesis in Drosophila melanogaster. Curr Biol 24:2673-80
Garaulet, Daniel L; Castellanos, Monica C; Bejarano, Fernando et al. (2014) Homeotic function of Drosophila Bithorax-complex miRNAs mediates fertility by restricting multiple Hox genes and TALE cofactors in the CNS. Dev Cell 29:635-48
Dai, Qi; Andreu-Agullo, Celia; Insolera, Ryan et al. (2013) BEND6 is a nuclear antagonist of Notch signaling during self-renewal of neural stem cells. Development 140:1892-902
Dai, Qi; Ren, Aiming; Westholm, Jakub O et al. (2013) The BEN domain is a novel sequence-specific DNA-binding domain conserved in neural transcriptional repressors. Genes Dev 27:602-14