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Related Concept Videos

Heterochromatin02:38

Heterochromatin

The extent of chromatin compaction can be studied by staining chromatin using specific DNA binding dyes. Under the microscope, the dense-compacted regions that take up more dye are called heterochromatin. Heterochromatin is further classified into two forms – constitutive heterochromatin and facultative heterochromatin.
Constitutive heterochromatin: It is a highly compact region of chromatin that is mostly concentrated in the centromere and telomere. Unlike euchromatin, the amino acid at 9th...
Histone Variants at the Centromere02:30

Histone Variants at the Centromere

Histone variants are the histone proteins with structural and sequence variations. These variants may be regarded as “mutant” forms that replace their canonical histone counterparts in the nucleosomes. Specific post-translational modifications on the histone variants enable further chromatin complexity and regulate tissue-specific gene expression. The most common histone variants are from histone H2A, H2B, and linker histone H1 families. However, several variants of histone H3 variants are also...
The Nucleosome Core Particle01:12

The Nucleosome Core Particle

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.
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their primary aim is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. On the other hand, they must allow polymerase enzymes to access histone-bound DNA during...
The Nucleosome Core Particle02:10

The Nucleosome Core Particle

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.
The paradox
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their main responsibility is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. While on the other hand, they must allow polymerase enzymes to access DNA...
Nucleosome Remodeling02:54

Nucleosome Remodeling

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.
Nucleosome remodeling complex
Eukaryotic cells have specialized enzymes called ATP-dependent nucleosome remodeling enzymes. These enzymes...
The Nucleosome01:19

The Nucleosome

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.
In a chromosome, DNA is wound twice around a protein complex called a histone octamer core, which consists of 8 histone proteins. This...

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Related Experiment Video

Updated: May 7, 2026

In Situ Nucleosome Assembly for Single-Molecule Correlative Force and Fluorescence Microscopy
05:58

In Situ Nucleosome Assembly for Single-Molecule Correlative Force and Fluorescence Microscopy

Published on: September 6, 2024

Human CAF-1-dependent nucleosome assembly in a defined system.

Lyudmila Y Kadyrova1, Elena Rodriges Blanko, Farid A Kadyrov

  • 1Department of Biochemistry and Molecular Biology; Southern Illinois University School of Medicine; Carbondale, IL USA.

Cell Cycle (Georgetown, Tex.)
|September 17, 2013
PubMed
Summary

Researchers developed a new in vitro system to study replication-coupled nucleosome assembly. This system requires key proteins like CAF-1 and PCNA, aiding chromatin packaging during DNA replication.

Keywords:
ASF1CAF-1DNA polymerase deltaPCNARFCchromatinhistone chaperonenucleosome assemblyreplication

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Assembly of Nucleosomal Arrays from Recombinant Core Histones and Nucleosome Positioning DNA

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Related Experiment Videos

Last Updated: May 7, 2026

In Situ Nucleosome Assembly for Single-Molecule Correlative Force and Fluorescence Microscopy
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In Situ Nucleosome Assembly for Single-Molecule Correlative Force and Fluorescence Microscopy

Published on: September 6, 2024

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique
06:32

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

Published on: March 9, 2022

Assembly of Nucleosomal Arrays from Recombinant Core Histones and Nucleosome Positioning DNA
10:40

Assembly of Nucleosomal Arrays from Recombinant Core Histones and Nucleosome Positioning DNA

Published on: September 10, 2013

Area of Science:

  • Molecular Biology
  • Epigenetics
  • Chromatin Dynamics

Background:

  • Replication-coupled nucleosome assembly is crucial for packaging newly synthesized DNA into chromatin.
  • Histone chaperones (CAF-1, ASF1A), PCNA, and RFC are known to be important.
  • Understanding this process in vitro has been challenging due to a lack of defined systems.

Purpose of the Study:

  • To develop a defined in vitro system that faithfully recapitulates replication-coupled nucleosome assembly.
  • To identify the essential components required for this process.

Main Methods:

  • Development of a defined in vitro system using purified proteins and DNA.
  • Assaying nucleosome assembly in the presence and absence of specific factors.
  • Utilizing nascent DNA synthesized by replicative polymerase delta.

Main Results:

  • A defined system was established that assembles nucleosomal arrays.
  • Assembly is dependent on CAF-1, ASF1A-H3-H4, H2A-H2B, PCNA, RFC, NAP1L1, ATP, and strand breaks.
  • Loss of CAF-1 p48 subunit significantly impairs DNA packaging.
  • The system forms nucleosomes on nascent DNA synthesized by polymerase delta.

Conclusions:

  • The developed system accurately reproduces key features of in vivo replication-coupled nucleosome assembly.
  • This system provides a valuable tool for further mechanistic studies of chromatin assembly during DNA replication.