Year: 2017
Type: article
Abstract: DNA adenine methyltransferase identification (DamID) is an enzymatic technology for detecting DNA regions targeted by chromatin-associated proteins. Proteins are fused to bacterial DNA adenine methyltransferase (Dam) and expressed in cultured cells or whole organisms. Here, we used DamID to detect DNA regions bound by the cardiac-restricted transcription factors (TFs) NKX2-5 and SRF, and ubiquitously-expressed co-factors ELK1 and ELK4. We compared targets bound by these TFs as N- and C-terminal fusions with Dam, for both wild type (WT) NKX2-5 and mutant proteins mimicking those found in congenital heart disease. Overall, DamID is highly robust: while the orientation of WT Dam fusions can affect the size of the target sets, their signatures remained largely reproducible. Furthermore, a severe NKX2-5 mutant lacking the homeodomain showed strong steric effects negatively impacting target discovery. The extent of steric effect is likely to be dependent on the protein in question and the orientation of Dam fusion.
Source: Genomics
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $2,980
Funders: National Science Foundation, Atlantic Philanthropies, Australian Government, Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung, University of New South Wales, State Government of Victoria, Medical Research Council, Australian Research Council, National Health and Medical Research Council
Awards: Gene regulatory networks in heart development, Genome-wide analysis of gene regulatory networks in heart development and congenital heart disease, G2742345854, Dynamics of the Nuclear Pore Proteins and the Mechanism of Transport, Probing the Cardiac Gene Regulatory Network in Development and Congenital Heart Disease, Australia Fellowship, Old genes learning new tricks: characterising regulatory changes driving increased heart complexity during vertebrate evolution, Molecular Approaches to Cardiac Development, Disease and Regeneration, Genetic pathways in heart development and congential heart disease, Unravelling gene networks in heart development and congenital heart disease, Transcriptional control of lineage specification in the developing heart
OpenAlex ID: W2586217554