抄録
Chromatin dynamics and higher order structures play essential roles in genomic DNA functions. Histone variants and histone post-translational modifications are involved in the regulation of chromatin structure and dynamics, cooperatively with DNA methylation and chromatin binding proteins. Therefore, studies of higher-order chromatin conformations have become important to reveal how genomic DNA is regulated during DNA transcription, replication, recombination and repair. The sedimentation velocity analysis by analytical ultracentrifugation has been commonly used to evaluate the higher-order conformation of in vitro reconstituted polynucleosomes, as model chromatin. Three major preparation methods for the unpurified, purified, and partially purified polynucleosomes have been reported so far. It is important to clarify the effects of the different polynucleosome preparation methods on the sedimentation profiles. To accomplish this, in the present study, we prepared unpurified, purified and partially purified polynucleosomes, and compared their sedimentation velocity profiles by analytical ultracentrifugation. In addition, we tested how the histone occupancy affects the sedimentation velocities of polynucleosomes. Our results revealed how free histones and polynucleosome aggregates affect the sedimentation velocity profiles of the polynucleosomes, in the absence and presence of Mg2+ ions.
| 元の言語 | English |
|---|---|
| ページ（範囲） | 381-388 |
| ページ数 | 8 |
| ジャーナル | Journal of Biochemistry |
| 巻 | 161 |
| 発行部数 | 4 |
| DOI | |
| 出版物ステータス | Published - 2017 4 1 |
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ASJC Scopus subject areas
- Medicine(all)
- Biochemistry
- Molecular Biology
これを引用
Influence of polynucleosome preparation methods on sedimentation velocity analysis of chromatin. / Kujirai, Tomoya; Machida, Shinichi; Osakabe, Akihisa; Kurumizaka, Hitoshi.
：: Journal of Biochemistry, 巻 161, 番号 4, 01.04.2017, p. 381-388.研究成果: Article
}
TY - JOUR
T1 - Influence of polynucleosome preparation methods on sedimentation velocity analysis of chromatin
AU - Kujirai, Tomoya
AU - Machida, Shinichi
AU - Osakabe, Akihisa
AU - Kurumizaka, Hitoshi
PY - 2017/4/1
Y1 - 2017/4/1
N2 - Chromatin dynamics and higher order structures play essential roles in genomic DNA functions. Histone variants and histone post-translational modifications are involved in the regulation of chromatin structure and dynamics, cooperatively with DNA methylation and chromatin binding proteins. Therefore, studies of higher-order chromatin conformations have become important to reveal how genomic DNA is regulated during DNA transcription, replication, recombination and repair. The sedimentation velocity analysis by analytical ultracentrifugation has been commonly used to evaluate the higher-order conformation of in vitro reconstituted polynucleosomes, as model chromatin. Three major preparation methods for the unpurified, purified, and partially purified polynucleosomes have been reported so far. It is important to clarify the effects of the different polynucleosome preparation methods on the sedimentation profiles. To accomplish this, in the present study, we prepared unpurified, purified and partially purified polynucleosomes, and compared their sedimentation velocity profiles by analytical ultracentrifugation. In addition, we tested how the histone occupancy affects the sedimentation velocities of polynucleosomes. Our results revealed how free histones and polynucleosome aggregates affect the sedimentation velocity profiles of the polynucleosomes, in the absence and presence of Mg2+ ions.
AB - Chromatin dynamics and higher order structures play essential roles in genomic DNA functions. Histone variants and histone post-translational modifications are involved in the regulation of chromatin structure and dynamics, cooperatively with DNA methylation and chromatin binding proteins. Therefore, studies of higher-order chromatin conformations have become important to reveal how genomic DNA is regulated during DNA transcription, replication, recombination and repair. The sedimentation velocity analysis by analytical ultracentrifugation has been commonly used to evaluate the higher-order conformation of in vitro reconstituted polynucleosomes, as model chromatin. Three major preparation methods for the unpurified, purified, and partially purified polynucleosomes have been reported so far. It is important to clarify the effects of the different polynucleosome preparation methods on the sedimentation profiles. To accomplish this, in the present study, we prepared unpurified, purified and partially purified polynucleosomes, and compared their sedimentation velocity profiles by analytical ultracentrifugation. In addition, we tested how the histone occupancy affects the sedimentation velocities of polynucleosomes. Our results revealed how free histones and polynucleosome aggregates affect the sedimentation velocity profiles of the polynucleosomes, in the absence and presence of Mg2+ ions.
KW - analytical ultracentrifugation
KW - chromatin
KW - nucleosome
KW - polynucleosome
KW - sedimentation velocity
UR - http://www.scopus.com/inward/record.url?scp=85021850339&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85021850339&partnerID=8YFLogxK
U2 - 10.1093/jb/mvw081
DO - 10.1093/jb/mvw081
M3 - Article
C2 - 28338761
AN - SCOPUS:85021850339
VL - 161
SP - 381
EP - 388
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 4
ER -