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Methods for Preparing Nucleosomes Containing Histone Variants

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1832)

Abstract

Histone variants are key epigenetic players that regulate transcription, repair, replication, and recombination of genomic DNA. Histone variant incorporation into nucleosomes induces structural diversity of nucleosomes, consequently leading to the structural versatility of chromatin. Such chromatin diversity created by histone variants may play a central role in the epigenetic regulation of genes. Each histone variant possesses specific biochemical and physical characteristics, and thus the preparation methods are complicated. Here, we introduce the methods for the purification of human histone variants as recombinant proteins, and describe the preparation methods for histone complexes and nucleosomes containing various histone variants. We also describe the detailed method for the preparation of heterotypic nucleosomes, which may function in certain biological phenomena. These methods are useful for biochemical, structural, and biophysical studies.

Key words

Histone variants Recombinant histones Nucleosome reconstitution Chromatin Histone 
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Notes

Acknowledgments

We thank Ms. Yukari Iikura (Waseda University) for her assistance. This work was supported in part by JSPS KAKENHI Grant Numbers JP25116002 and JP17H01408, by Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)) from AMED under Grant Number JP18am0101076, and by JST CREST Grant Number PMJCR16G1, Japan [to H.K.]. H.K. and N.H. were supported by the Waseda Research Institute for Science and Engineering. T.K. and R.F. were supported by Research Fellowships from JSPS for Young Scientists [JP15J06807 and JP16J10043, respectively]. Y.A. and S.M. were supported in part by JSPS KAKENHI Grant Numbers JP17K15043 [to Y.A.] and JP16K18473 [to S.M.].

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory of Chromatin Structure and Function, Institute for Quantitative BiosciencesThe University of TokyoBunkyo-kuJapan
  2. 2.Graduate School of Advanced Science and EngineeringWaseda UniversityShinjuku-kuJapan

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