TY - JOUR
T1 - Histone H3.5 forms an unstable nucleosome and accumulates around transcription start sites in human testis
AU - Urahama, Takashi
AU - Harada, Akihito
AU - Maehara, Kazumitsu
AU - Horikoshi, Naoki
AU - Sato, Koichi
AU - Sato, Yuko
AU - Shiraishi, Koji
AU - Sugino, Norihiro
AU - Osakabe, Akihisa
AU - Tachiwana, Hiroaki
AU - Kagawa, Wataru
AU - Kimura, Hiroshi
AU - Ohkawa, Yasuyuki
AU - Kurumizaka, Hitoshi
N1 - Funding Information:
We thank Takako Ichinose (Kyushu University) for the ChIP-Seq analysis and Yasuhiro Arimura (Waseda University) for discussions. The synchrotron radiation experiments were performed at the BL41XU beamline of SPring-8, with the approval of the Japan Synchrotron Radiation Research Institute (JASRI) (Proposal No. 2011A1528). We are grateful to the beamline scientists for their assistance in data collection. This work was supported by MEXT KAKENHI (25116002 to H.Kurumizaka, 25116005 to H.Kimura, 25131706 to Y.O., and 26116521 to W.K.) and the Platform Project for Supporting Drug Discovery and Life Science Research (Platform for Drug Discovery, Informatics, and Structural Life Science) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) and Japan Agency for Medical Research and Development (AMED) to H.Kurumizaka. H.Kurumizaka and N.H. were supported by the Waseda Research Institute for Science and Engineering and Waseda University. T.U. was supported by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists.
Publisher Copyright:
© 2016 Urahama et al.
PY - 2016/1/15
Y1 - 2016/1/15
N2 - Background: Human histone H3.5 is a non-allelic H3 variant evolutionally derived from H3.3. The H3.5 mRNA is highly expressed in human testis. However, the function of H3.5 has remained poorly understood. Results: We found that the H3.5 nucleosome is less stable than the H3.3 nucleosome. The crystal structure of the H3.5 nucleosome showed that the H3.5-specific Leu103 residue, which corresponds to the H3.3 Phe104 residue, reduces the hydrophobic interaction with histone H4. Mutational analyses revealed that the H3.5-specific Leu103 residue is responsible for the instability of the H3.5 nucleosome, both in vitro and in living cells. The H3.5 protein was present in human seminiferous tubules, but little to none was found in mature sperm. A chromatin immunoprecipitation coupled with sequencing analysis revealed that H3.5 accumulated around transcription start sites (TSSs) in testicular cells. Conclusions: We performed comprehensive studies of H3.5, and found the instability of the H3.5 nucleosome and the accumulation of H3.5 protein around TSSs in human testis. The unstable H3.5 nucleosome may function in the chromatin dynamics around the TSSs, during spermatogenesis.
AB - Background: Human histone H3.5 is a non-allelic H3 variant evolutionally derived from H3.3. The H3.5 mRNA is highly expressed in human testis. However, the function of H3.5 has remained poorly understood. Results: We found that the H3.5 nucleosome is less stable than the H3.3 nucleosome. The crystal structure of the H3.5 nucleosome showed that the H3.5-specific Leu103 residue, which corresponds to the H3.3 Phe104 residue, reduces the hydrophobic interaction with histone H4. Mutational analyses revealed that the H3.5-specific Leu103 residue is responsible for the instability of the H3.5 nucleosome, both in vitro and in living cells. The H3.5 protein was present in human seminiferous tubules, but little to none was found in mature sperm. A chromatin immunoprecipitation coupled with sequencing analysis revealed that H3.5 accumulated around transcription start sites (TSSs) in testicular cells. Conclusions: We performed comprehensive studies of H3.5, and found the instability of the H3.5 nucleosome and the accumulation of H3.5 protein around TSSs in human testis. The unstable H3.5 nucleosome may function in the chromatin dynamics around the TSSs, during spermatogenesis.
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U2 - 10.1186/s13072-016-0051-y
DO - 10.1186/s13072-016-0051-y
M3 - Article
AN - SCOPUS:84954476360
VL - 9
JO - Epigenetics and Chromatin
JF - Epigenetics and Chromatin
SN - 1756-8935
IS - 1
M1 - 51
ER -