000114904 001__ 114904
000114904 005__ 20190316234108.0
000114904 0247_ $$2doi$$a10.1093/nar/gni079
000114904 02470 $$2ISI$$a000229113100010
000114904 037__ $$aARTICLE
000114904 245__ $$aProof of concept for microarray-based detection of DNA-binding oncogenes in cell extracts
000114904 269__ $$a2005
000114904 260__ $$c2005
000114904 336__ $$aJournal Articles
000114904 520__ $$aThe function of DNA-binding proteins is controlled not just by their abundance, but mainly at the level of their activity in terms of their interactions with DNA and protein targets. Moreover, the affinity of such transcription factors to their target sequences is often controlled by co-factors and/or modifications that are not easily assessed from biological samples. Here, we describe a scalable method for monitoring protein-DNA interactions on a microarray surface. This approach was designed to determine the DNA-binding activity of proteins in crude cell extracts, complementing conventional expression profiling arrays. Enzymatic labeling of DNA enables direct normalization of the protein binding to the microarray, allowing the estimation of relative binding affinities. Using DNA sequences covering a range of affinities, we show that the new microarray-based method yields binding strength estimates similar to low-throughput gel mobility-shift assays. The microarray is also of high sensitivity, as it allows the detection of a rare DNA-binding protein from breast cancer cells, the human tumor suppressor AP-2. This approach thus mediates precise and robust assessment of the activity of DNA-binding proteins and takes present DNA-binding assays to a high throughput level.
000114904 700__ $$aEgener, T.
000114904 700__ $$aRoulet, E.
000114904 700__ $$aZehnder, M.
000114904 700__ $$0244404$$g113607$$aBucher, P.
000114904 700__ $$aMermod, N.
000114904 773__ $$j33$$tNucleic Acids Res$$k8$$qe79
000114904 8564_ $$uhttps://infoscience.epfl.ch/record/114904/files/Nucl.%20Acids%20Res.-2005-Egener-e79.pdf$$zPublisher's version$$s314813$$yPublisher's version
000114904 909C0 $$xU11780$$0252244$$pGR-BUCHER
000114904 909CO $$ooai:infoscience.tind.io:114904$$qGLOBAL_SET$$pSV$$particle
000114904 917Z8 $$x182396
000114904 937__ $$aGR-BUCHER-ARTICLE-2005-001
000114904 973__ $$rREVIEWED$$sPUBLISHED$$aOTHER
000114904 980__ $$aARTICLE