Related Experiment Video
Updated: Apr 15, 2026
08:16
In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays
Published on: December 29, 2021
3.3K
Methods to study histone chaperone function in nucleosome assembly and chromatin transcription
Parijat Senapati1, Deepthi Sudarshan, Shrikanth S Gadad
1Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur PO, Bangalore, Karnataka, 560064, India.
Methods in Molecular Biology (Clifton, N.J.)
|April 2, 2015
Summary
Histone chaperones manage histone proteins for DNA processes. This chapter details methods to study their role in transcription, focusing on human Nucleophosmin (NPM1).
Area of Science:
- Molecular Biology
- Epigenetics
- Biochemistry
Background:
- Histone chaperones are crucial proteins regulating histone dynamics.
- These proteins are vital for DNA-templated processes like replication, repair, and transcription.
- Histone chaperone complexes, such as FACT, are implicated in nucleosome dynamics during transcription.
Purpose of the Study:
- To describe methods for studying histone chaperone activity.
- To elucidate the role of histone chaperones in transcription.
- To highlight the function of human Nucleophosmin (NPM1) as a transcription co-activator.
Main Methods:
- Histone chaperone activity assays.
- Techniques for studying protein-nucleosome interactions.
- Methods to assess the impact of histone chaperones on transcription.
Main Results:
- Established methods for analyzing histone chaperone functions.
- Demonstrated the involvement of histone chaperones in transcriptional regulation.
- Confirmed human Nucleophosmin (NPM1) as a transcription co-activator.
Conclusions:
- Histone chaperones are key regulators of transcription.
- The described methods provide a framework for investigating histone chaperone roles.
- Human Nucleophosmin (NPM1) plays a significant role in co-activating transcription.
Related Concept Videos
02:54Nucleosome Remodeling
11.7K
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...
Nucleosome remodeling complex
Eukaryotic cells have specialized enzymes called ATP-dependent nucleosome remodeling enzymes. These enzymes...
11.7K
02:05Duplication of Chromatin Structure
7.7K
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...
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...
7.7K
01:12The Nucleosome Core Particle
2.8K
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...
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...
2.8K
02:10The Nucleosome Core Particle
15.4K
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...
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...
15.4K
02:25Spreading of Chromatin Modifications
10.1K
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...
Writers
The writer...
10.1K
02:36Chromatin Immunoprecipitation- ChIP
12.9K
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.
Types of ChIP
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...
Types of ChIP
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...
12.9K