Abstract

The majority of histones are replaced by protamines during spermatogenesis, but small amounts are retained in mammalian spermatozoa. Since nucleosomes in spermatozoa influence epigenetic inheritance, it is important to know how histones are distributed in the sperm genome. Conflicting data, which may result from different conditions used for micrococcal nuclease (MNase) digestion, have been reported: retention of nucleosomes at either gene promoter regions or within distal gene-poor regions. Here, we find that the swim-up sperm used in many studies contain about 10% population of sperm which have not yet completed the histone-to-protamine replacement. We develop a method to purify histone replacement-completed sperm (HRCS) and to completely solubilize histones from cross-linked HRCS without MNase digestion. Our results indicate that histones are retained at specific promoter regions in HRCS. This method allows the study of epigenetic status in mature sperm.

Details

Title
Mapping of histone-binding sites in histone replacement-completed spermatozoa
Author
Yoshida, Keisuke 1   VIAFID ORCID Logo  ; Muratani, Masafumi 2   VIAFID ORCID Logo  ; Araki, Hiromitsu 3 ; Miura, Fumihito 3 ; Suzuki, Takehiro 4 ; Dohmae, Naoshi 4 ; Katou, Yuki 5 ; Shirahige, Katsuhiko 5 ; Ito, Takashi 3 ; Ishii, Shunsuke 6 

1  Cluster for Pioneering Research, CREST Research Project of JST (Japan Science and Technology Agency), RIKEN Tsukuba Institute, Tsukuba, Ibaraki, Japan 
2  Department of Genome Biology, Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki, Japan 
3  Department of Biochemistry, Kyushu University Graduate School of Medical Sciences, Fukuoka, Japan 
4  Biomolecular Characterization Unit, RIKEN Center for Sustainable Resource Science, Wako, Saitama, Japan 
5  Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan 
6  Cluster for Pioneering Research, CREST Research Project of JST (Japan Science and Technology Agency), RIKEN Tsukuba Institute, Tsukuba, Ibaraki, Japan; Department of Functional Genomics, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan 
Pages
1-11
Publication year
2018
Publication date
Sep 2018
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-018-06243-9
ProQuest document ID
2111724353
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.