The BEN domain is a novel sequence-specific DNA-binding domain conserved in neural transcriptional repressors Journal Article

   

Authors:	Dai, Q.; Ren, A.; Westholm, J. O.; Serganov, A. A.; Patel, D. J.; Lai, E. C.
Article Title:	The BEN domain is a novel sequence-specific DNA-binding domain conserved in neural transcriptional repressors
Abstract:	We recently reported that Drosophila Insensitive (Insv) promotes sensory organ development and has activity as a nuclear corepressor for the Notch transcription factor Suppressor of Hairless [Su(H)]. Insv lacks domains of known biochemical function but contains a single BEN domain (i.e., a "BEN-solo" protein). Our chromatin immunoprecipitation (ChIP) sequencing (ChIP-seq) analysis confirmed binding of Insensitive to Su(H) target genes in the Enhancer of split gene complex [E(spl)-C]; however, de novo motif analysis revealed a novel site strongly enriched in Insv peaks (TCYAATHRGAA). We validate binding of endogenous Insv to genomic regions bearing such sites, whose associated genes are enriched for neural functions and are functionally repressed by Insv. Unexpectedly, we found that the Insv BEN domain binds specifically to this sequence motif and that Insv directly regulates transcription via this motif. We determined the crystal structure of the BEN-DNA target complex, revealing homodimeric binding of the BEN domain and extensive nucleotide contacts via α helices and a C-terminal loop. Point mutations in key DNA-contacting residues severely impair DNA binding in vitro and capacity for transcriptional regulation in vivo. We further demonstrate DNA-binding and repression activities by the mammalian neural BEN-solo protein BEND5. Altogether, we define novel DNA-binding activity in a conserved family of transcriptional repressors, opening a molecular window on this extensive gene family. © 2013 by Cold Spring Harbor Laboratory Press.
Keywords:	controlled study; unclassified drug; dna binding protein; sequence analysis; nonhuman; protein domain; protein function; protein motif; animal cell; gene targeting; complex formation; gene expression; carboxy terminal sequence; animal experiment; protein binding; transcription factor; drosophila; in vivo study; in vitro study; dna; transcription regulation; chromatin immunoprecipitation; gene repression; binding protein; binding site; crystal structure; point mutation; dna binding; repressor protein; neurogenesis; nerve cell; genomic dna; drosophila protein; nucleotide; repressor gene; alpha helix; transcriptional repression; homodimer; ben domain; ben protein; bend5 protein; enhancer of split protein complex; insv protein; suh protein
Journal Title:	Genes and Development
Volume:	27
Issue:	6
ISSN:	0890-9369
Publisher:	Cold Spring Harbor Laboratory Press
Date Published:	2013-03-15
Start Page:	602
End Page:	614
Language:	English
PROVIDER:	scopus
PMCID:	PMC3613608
PUBMED:	23468431
DOI:	10.1101/gad.213314.113
DOI/URL:	http://www.scopus.com/inward/record.url?eid=2-s2.0-84875328376&partnerID=40&md5=f74bb785ec293f9c733c220fcefa8db6
Notes:	--- - "Export Date: 1 May 2013" - "CODEN: GEDEE" - ":doi 10.1101/gad.213314.113" - ": Chemicals/CASDNA, 9007-49-2" - "Source: Scopus"

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