ArticleProceedings of the National Academy of Sciences of the United States of America2012
Quantifying transient binding of ISWI chromatin remodelers in living cells by pixel-wise photobleaching profile evolution analysis.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 12 citations in OpenAlex.
- DeepFRAP: Fast fluorescence recovery after photobleaching data analysis using deep neural networks.Journal of microscopy · 2021Article
- IMITATION SWITCH is required for normal chromatin structure and gene repression in PRC2 target domains.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- A Highly Accurate Pixel-Based FRAP Model Based on Spectral-Domain Numerical Methods.Biophysical journal · 2019Article
- A Discontinuous Galerkin Model for Fluorescence Loss in Photobleaching.Scientific reports · 2018Article
- Quantitative imaging of mammalian transcriptional dynamics: from single cells to whole embryos.BMC biology · 2016Review
- Active promoters give rise to false positive 'Phantom Peaks' in ChIP-seq experiments.Nucleic acids research · 2015Article
- Second German-Catalan workshop on epigenetics & cancer.Epigenetics · 2015Article
- Retrieving the intracellular topology from multi-scale protein mobility mapping in living cells.Nature communications · 2014Article
- Molecular diffusion and binding analyzed with FRAP.Protoplasma · 2014Review
- Advanced fluorescence microscopy methods for the real-time study of transcription and chromatin dynamics.Transcription · 2014Review
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2 authors at 2 institutions in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
Interactions between nuclear proteins and chromatin frequently occur on the time scale of seconds and below. These transient binding events are important for the fast identification of target sites as concluded from our previous analysis of the human chromatin remodelers Snf2H and Snf2L from the imitation switch (ISWI) family. Both ATP-driven molecular motor proteins are able to translocate nucleosomes along the DNA and appear to exert this activity only on a small number of nucleosomes to which they bind more tightly. For mechanistic studies, one needs to distinguish such translocation reactions or other long-lived interactions associated with conformational changes and/or ATP hydrolysis from nonproductive chromatin sampling during target search. These processes can be separated by measuring the duration of nucleosome binding with subsecond time resolution. To reach this goal, we have developed a fluorescence bleaching technique termed pixel-wise photobleaching profile evolution analysis (3PEA). It exploits the inherent time structure of confocal microscopy images and yields millisecond resolution. 3PEA represents a generally applicable approach to quantitate transient chromatin interactions in the 2- to 500-ms time regime within only ∼1 s needed for a measurement. The green autofluorescent protein (GFP)-tagged Snf2H and Snf2L and the inactive Snf2L+13 splice variant were studied by 3PEA in comparison to the isolated GFP or red autofluorescent protein and a GFP pentamer. Our results reveal that the residence time for transient chromatin binding of Snf2H and Snf2L is <2 ms, and strongly support the view that ISWI-type remodelers are only rarely active in unperturbed cells during G1 phase.
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