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Related Concept Videos

Chromatin Packaging02:21

Chromatin Packaging

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Each human somatic cell contains 6 billion base-pairs of DNA. Each base-pair is 0.34 nm long, which means that each diploid cell contains a staggering 2 meters of DNA. How is such a long DNA strand packed inside a nucleus measuring only 10 - 20 microns in diameter? 
The chromatin
In combination with specialized DNA binding protein called Histones, the DNA double helix forms a compact DNA: protein complex called chromatin. The chromatin itself is further compacted into higher-order...
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Spreading of Chromatin Modifications02:25

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The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
Writers
The writer...
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Chromatin Immunoprecipitation- ChIP02:36

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Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
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ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
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Inheritance of Chromatin Structures03:17

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Epigenetics is the study of inherited changes in a cell's phenotype without changing the DNA sequences. It provides a form of memory for the differential gene expression pattern to maintain cell lineage, position-effect variegation, dosage compensation, and maintenance of chromatin structures such as telomeres and centromeres. For example, the structure and location of the centromere on chromosomes are epigenetically inherited. Its functionality is not dictated or ensured by the underlying...
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Duplication of Chromatin Structure02:05

Duplication of Chromatin Structure

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The process of chromosome duplication during cell division requires genome-wide disruption and re-assembly of chromatin. The chromatin structure must be accurately inherited, reassembled, and maintained in the daughter cells to ensure lineage propagation.
The basic unit of the chromatin is the nucleosome, consisting of DNA wrapped around octameric histone proteins and short stretches of linker DNA separating individual nucleosomes. The histone proteins within the nucleosome have their...
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Polytene Chromosomes02:04

Polytene Chromosomes

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Polytene chromosomes are giant interphase chromosomes with several DNA strands placed side by side. They were discovered in the year 1881 by Balbiani in salivary glands, intestine, muscles, malpighian tubules, and hypoderm of larvae Chironomus plumosus. Hence, these are also called "Salivary gland chromosomes." These are found in insects of the order Diptera and Collembola; in certain organs of mammals; and synergids, antipodes of flowering plants. Polytene chromosomes are also...
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Related Experiment Video

Updated: Jul 23, 2025

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

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

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DNA curtains to visualize chromatin interactions.

Mitchell Woodhouse1, J Brooks Crickard1

  • 1Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.

Methods (San Diego, Calif.)
|July 12, 2023
PubMed
Summary
This summary is machine-generated.

Visualize protein-DNA interactions in real-time using DNA curtains and total internal reflection microscopy (TIRFM). This method allows direct observation of enzyme function on chromatinized DNA, advancing our understanding of DNA-related biological processes.

Keywords:
Fluorescent microscopyHistonesNucleosomeSingle molecule

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Area of Science:

  • Biochemistry and Molecular Biology
  • Microscopy and Imaging Techniques

Background:

  • Single-molecule imaging has revolutionized the study of dynamic biochemical processes.
  • Total internal reflection microscopy (TIRFM) enables real-time visualization of protein-DNA interactions.
  • Existing techniques have enhanced understanding of DNA transcription, replication, repair, and recombination within chromatin.

Purpose of the Study:

  • To introduce a general single-molecule technique, DNA curtains, for visualizing enzyme function on chromatinized DNA.
  • To provide methods for expressing and reconstituting nucleosomes on long DNA stretches.
  • To demonstrate direct visualization of these reconstituted nucleosomes using TIRFM and DNA curtains.

Main Methods:

  • Development and application of the DNA curtains technique.
  • Utilizing total internal reflection microscopy (TIRFM) for high-resolution imaging.
  • Methods for nucleosome expression and reconstitution on extended DNA molecules.

Main Results:

  • Demonstration of a robust method for visualizing enzyme activity on chromatinized DNA in real-time.
  • Successful reconstitution of nucleosomes on long DNA fragments suitable for single-molecule analysis.
  • Direct visualization of protein-DNA interactions within a chromatin context using TIRFM.

Conclusions:

  • The DNA curtains technique offers a powerful platform for studying enzyme mechanisms on chromatin.
  • This approach significantly advances the ability to observe complex DNA-protein interactions directly.
  • The described methods facilitate deeper insights into fundamental DNA processes like repair and replication.