1. WDR5, BRCA1, and BARD1 Co-regulate the DNA Damage Response and Modulate the Mesenchymal-to-Epithelial Transition during Early Reprogramming.
- Author
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Peñalosa-Ruiz G, Bousgouni V, Gerlach JP, Waarlo S, van de Ven JV, Veenstra TE, Silva JCR, van Heeringen SJ, Bakal C, Mulder KW, and Veenstra GJC
- Subjects
- Animals, BRCA1 Protein metabolism, Chromatin genetics, Chromatin metabolism, DNA Repair, Gene Expression Profiling, Humans, Intracellular Signaling Peptides and Proteins metabolism, Mice, Phenotype, Signal Transduction, Transcription Factors genetics, Transcription Factors metabolism, Tumor Suppressor Proteins metabolism, Ubiquitin-Protein Ligases metabolism, BRCA1 Protein genetics, Cellular Reprogramming genetics, DNA Damage, Epithelial-Mesenchymal Transition genetics, Intracellular Signaling Peptides and Proteins genetics, Tumor Suppressor Proteins genetics, Ubiquitin-Protein Ligases genetics
- Abstract
Differentiated cells are epigenetically stable, but can be reprogrammed to pluripotency by expression of the OSKM transcription factors. Despite significant effort, relatively little is known about the cellular requirements for reprogramming and how they affect the properties of induced pluripotent stem cells. We have performed high-content screening with small interfering RNAs targeting 300 chromatin-associated factors and extracted colony-level quantitative features. This revealed five morphological phenotypes in early reprogramming, including one displaying large round colonies exhibiting an early block of reprogramming. Using RNA sequencing, we identified transcriptional changes associated with these phenotypes. Furthermore, double knockdown epistasis experiments revealed that BRCA1, BARD1, and WDR5 functionally interact and are required for the DNA damage response. In addition, the mesenchymal-to-epithelial transition is affected in Brca1, Bard1, and Wdr5 knockdowns. Our data provide a resource of chromatin-associated factors in early reprogramming and underline colony morphology as an important high-dimensional readout for reprogramming quality., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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