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Chromatin Modification in iPS Cells01:32

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Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
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Visualizing, quantifying and mapping chromatin remodelers at work with single-molecule and single-cell imaging.

Hendrik Sielaff1, Ziqing Winston Zhao2

  • 1Department of Chemistry, Faculty of Science, National University of Singapore, Singapore 119543, Singapore; Centre for BioImaging Sciences (CBIS), Faculty of Science, National University of Singapore, Singapore 117557, Singapore.

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Recent advances in imaging reveal how chromatin remodelers function at the molecular level. These studies illuminate the dynamics and organization of these complexes, offering insights into genome regulation and disease.

Keywords:
Chromatin remodelingDNA-binding dynamicsNucleosome translocationPhase separationSingle-molecule imaging

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • Cell Biology

Background:

  • Chromatin remodeling is essential for regulating genome access by overcoming nucleosome-imposed topological challenges.
  • ATP-dependent remodeler complexes play a crucial role in this process across eukaryotes.
  • Understanding the dynamics and mechanisms of chromatin remodelers is vital for comprehending gene regulation.

Purpose of the Study:

  • To provide an integrated overview of recent advancements in visualizing and quantifying chromatin remodeler activity.
  • To elucidate the spatial and temporal aspects of chromatin remodeling processes.
  • To explore the functional coupling of chromatin remodelers with transcription and their role in disease.

Main Methods:

  • Application of single-molecule and single-cell imaging techniques, both in vitro and in cellulo.
  • Quantitative analysis of chromatin remodeler dynamics, binding, and target search.
  • Mapping the intranuclear organization of remodelers, including hotspots and phase condensates.

Main Results:

  • Visualization and quantification of molecular mechanisms like DNA wrapping/unwrapping, nucleosome translocation, and histone exchange.
  • Detailed insights into the dynamics of chromatin binding and target search by remodelers.
  • Characterization of intranuclear organization into hotspots and phase condensates, and coupling with transcription.

Conclusions:

  • Recent imaging techniques provide unprecedented mechanistic insights into chromatin remodeling across molecular and cellular scales.
  • Quantitative parameters reveal a multi-modal regulatory landscape for chromatin remodelers.
  • Further research into misregulation of chromatin remodeling is crucial for understanding disease contexts.