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Related Concept Videos

The Nucleosome02:33

The Nucleosome

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DNA in a human cell is almost 2m long and it is packed inside a tiny nucleus that is only a few microns in diameter. The level of compaction of DNA inside the nucleus is astonishing. It is organized into several sequentially higher levels of compaction to fit into such a tiny space. The most compact form of DNA is a chromosome that can be seen under a microscope in a dividing cell.
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The Nucleosome01:19

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Human DNA is almost two meters long. However, it is compressed inside a tiny nucleus measuring only a few microns in diameter. To make this degree of compaction possible, DNA is organized into several sequential levels so that it can fit into such a tiny space. The most compact form of DNA is a chromosome that can be seen under a microscope in a dividing cell.
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Nucleosomes are the basic units of chromatin compaction. Each nucleosome consists of the DNA bound tightly around a histone core, which makes the DNA inaccessible to DNA binding proteins such as DNA polymerase and RNA polymerase. Hence, the fundamental problem is to ensure access to DNA when appropriate, despite the compact and protective chromatin structure.
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Nucleosomes are the DNA-histone complex, where the DNA strand is wound around the histone core. The histone core is an octamer containing two copies of H2A, H2B, H3, and H4 histone proteins.
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Nucleosomes are the DNA-histone complex, where the DNA strand is wound around the histone core. The histone core is an octamer containing two copies of H2A, H2B, H3, and H4 histone proteins.
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Measuring genome-wide nucleosome turnover using CATCH-IT.

Sheila S Teves1, Roger B Deal, Steven Henikoff

  • 1Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA.

Methods in Enzymology
|August 30, 2012
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Summary

This study introduces Covalent Attachment of Tagged Histones to Capture and Identify Turnover (CATCH-IT), a genome-wide method to measure nucleosome turnover kinetics. This technique offers insights into dynamic chromatin regulation for essential nuclear processes.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Epigenetics

Background:

  • Nucleosome dynamics are crucial for fundamental nuclear processes like DNA transcription, replication, and repair.
  • Understanding nucleosome assembly, eviction, and replacement is key to regulating these dynamic chromatin processes.

Purpose of the Study:

  • To present a genome-wide method for measuring nucleosome turnover kinetics.
  • To provide a protocol for high-resolution mapping of nucleosome and subnucleosomal particles.

Main Methods:

  • Metabolic labeling of newly synthesized histones.
  • Capture of labeled histones using the CATCH-IT (Covalent Attachment of Tagged Histones to Capture and Identify Turnover) method.
  • High-resolution genome-wide profiling using paired-end sequencing and Solexa DNA library preparation.

Main Results:

  • The CATCH-IT method enables genome-wide measurement of nucleosome turnover.
  • High-resolution mapping of nucleosome and subnucleosomal particles is achievable with single base-pair resolution.
  • The protocol is demonstrated in a Drosophila cell line and adaptable to other model systems.

Conclusions:

  • Nucleosome turnover kinetics can be effectively measured using the CATCH-IT method.
  • This technique provides valuable insights into the regulation of dynamic chromatin processes.
  • The described protocols facilitate detailed analysis of chromatin structure and dynamics across different model organisms.