000125659 001__ 125659
000125659 005__ 20181203021249.0
000125659 0247_ $$2doi$$a10.1101/gr.2445504
000125659 037__ $$aARTICLE
000125659 245__ $$aA gateway-compatible yeast one-hybrid system
000125659 269__ $$a2004
000125659 260__ $$c2004
000125659 336__ $$aJournal Articles
000125659 500__ $$aHighlighted in: Nat Methods, 1, 190, 2004
000125659 520__ $$aSince the advent of microarrays, vast amounts of gene expression data have been generated. However, these microarray data fail to reveal the transcription regulatory mechanisms that underlie differential gene expression, because the identity of the responsible transcription factors (TFs) often cannot be directly inferred from such data sets. Regulatory TFs activate or repress transcription of their target genes by binding to cis-regulatory elements that are frequently located in a gene's promoter. To understand the mechanisms underlying differential gene expression, it is necessary to identify physical interactions between regulatory TFs and their target genes. We developed a Gateway-compatible yeast one-hybrid (Y1H) system that enables the rapid, large-scale identification of protein-DNA interactions using both small (i.e., DNA elements of interest) and large (i.e., gene promoters) DNA fragments. We used four well-characterized Caenorhabditis elegans promoters as DNA baits to test the functionality of this Y1H system. We could detect approximately 40% of previously reported TF-promoter interactions. By performing screens using two complementary libraries, we found novel potentially interacting TFs for each promoter. We recapitulated several of the Y1H-based protein-DNA interactions using luciferase reporter assays in mammalian cells. Taken together, the Gateway-compatible Y1H system will allow the high-throughput identification of protein-DNA interactions and may be a valuable tool to decipher transcription regulatory networks.
000125659 6531_ $$aAnimals
000125659 6531_ $$aCaenorhabditis elegans/physiology
000125659 6531_ $$aComputational Biology/methods
000125659 6531_ $$aDNA
000125659 6531_ $$aHelminth/chemistry/metabolism
000125659 6531_ $$aDNA-Binding Proteins/physiology
000125659 6531_ $$aGene Expression Regulation
000125659 6531_ $$aGenome
000125659 6531_ $$aLuciferases/metabolism
000125659 6531_ $$aPromoter Regions (Genetics)/genetics
000125659 6531_ $$aRegulatory Sequences
000125659 6531_ $$aNucleic Acid/physiology
000125659 6531_ $$aSaccharomyces cerevisiae/physiology
000125659 6531_ $$aTranscription Factors/isolation & purification/physiology
000125659 6531_ $$aTranscription
000125659 6531_ $$aGenetic
000125659 700__ $$0244717$$g182376$$aDeplancke, B.
000125659 700__ $$aDupuy, D.
000125659 700__ $$aVidal, M.
000125659 700__ $$aWalhout, A. J.
000125659 773__ $$j14$$tGenome Res$$k10B$$q2093-101
000125659 909C0 $$xU11829$$0252247$$pUPDEPLA
000125659 909CO $$pSV$$particle$$ooai:infoscience.tind.io:125659
000125659 937__ $$aUPDEPLA-ARTICLE-2004-002
000125659 973__ $$rREVIEWED$$sPUBLISHED$$aOTHER
000125659 980__ $$aARTICLE