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Mol. Hum. Reprod. Advance Access originally published online on April 3, 2009
Molecular Human Reproduction 2009 15(6):373-378; doi:10.1093/molehr/gap028
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© The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Mass spectrometry analysis of dynamic post-translational modifications of TH2B during spermatogenesis

Shuang Lu1,2, Yong M. Xie4, Xin Li2, Ji Luo5, Xin Q. Shi3, Xin Hong4, Ying H. Pan5 and Xu Ma1,2,6

1Graduate School, Peking Union Medical College, Beijing, China 2Genetics Department of the National Institute Research for Family Planning, WHO Human Reproduction Research Collaborative Center, Beijing, China 3 Endocrine Department of the National Institute Research for Family Planning, Beijing, China 4Bejing Office, Bruker Daltonics Inc., Beijing, China 5 Institute of Crop Science, Chinese Academy of Agricultural Sciences/The National Key Facility for Crop Gene Resources and Genetic Improvement, NFCRI, Beijing, China

6 Correspondence address. E-mail: genetic{at}263.net.cn

TH2B, an important testis histone, plays a key role in remodeling chromatin structure during spermatogenesis. We present a detailed study of post-translational modifications (PTMs) of histone TH2B from different developmental stages of sperm cells, using a combination of high performance liquid chromatography, enzymatic Glu-c digestions of peptides, liquid chromatography–mass spectrometry (LC–MS) and LC–MS/MS analysis. The results showed modification patterns of the intact histone TH2B during spermatogenesis. Acetylated TH2B was most abundant in spermatogonia (28.9%) when compared with the spermatocytes (8.3%) and round spermatids (11.2%). Several new PTMs of TH2B were identified. In spermatogonia, spermatocytes and round spermatids, T116 and K117, were modified by phosphorylation and methylation, respectively, forming a novel ‘phospho switch’ site. The identified modification patterns of histone TH2B in spermatogenic cells provides a basis for future studies on histone coding and epigenetic regulation during spermatogenesis.

Key words: spermatogenesis/post-translational modifications/chromatin/testis-specific histones/TH2B

Submitted on February 11, 2009; resubmitted on March 11, 2009; accepted on March 31, 2009.


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