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Related Experiment Video

Updated: Jun 7, 2026

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

Published on: March 3, 2016

Epigenetic inheritance of centromeres.

S Henikoff¹, T Furuyama

¹Howard Hughes Medical Institute, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA. steveh@fhcrc.org

Cold Spring Harbor Symposia on Quantitative Biology

|November 5, 2010

Summary

Centromere inheritance relies on unique histone variants (CenH3) forming nucleosomes that induce positive supercoiling. This distinct DNA wrapping mechanism ensures centromere identity without DNA sequence dependence.

Area of Science:

Epigenetics
Molecular Biology
Genetics

Background:

Centromeres are crucial for chromosome segregation but their epigenetic inheritance mechanism remains unclear.
Centromere identity depends on nucleosomes containing the histone variant CenH3 instead of canonical H3.
Selfish centromere drive during female meiosis may have driven the evolution of epigenetic centromere inheritance.

Purpose of the Study:

To investigate the mechanism of epigenetic centromere inheritance.
To understand the structural and functional properties of CenH3-containing nucleosomes.
To propose a model for how centromere identity is maintained.

Main Methods:

Molecular studies of Drosophila and yeast centromeres.
Analysis of nucleosome structure and DNA supercoiling.

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Analysis of Transgenerational Epigenetic Inheritance in C. elegans Using a Fluorescent Reporter and Chromatin Immunoprecipitation (ChIP)

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Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins

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Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

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Analysis of Transgenerational Epigenetic Inheritance in C. elegans Using a Fluorescent Reporter and Chromatin Immunoprecipitation (ChIP)

Analysis of Transgenerational Epigenetic Inheritance in C. elegans Using a Fluorescent Reporter and Chromatin Immunoprecipitation (ChIP)

Published on: May 5, 2023

In vivo functional assays.

Main Results:

Drosophila CenH3 nucleosomes are half the size of canonical H3 nucleosomes.
CenH3 nucleosomes induce positive supercoils, unlike H3 nucleosomes which induce negative supercoils.
Positive supercoiling suggests a right-handed DNA wrap, implying non-octameric structures and exposed surfaces for protein binding.

Conclusions:

The unique topology of CenH3 nucleosomes likely explains centromere stability and maintenance as unique loci.
This DNA wrapping mechanism provides a sequence-independent mode of centromere inheritance.
Incompatibility of opposing nucleosome topologies may prevent centromere drive and ensure faithful inheritance.