Lu Shuang, Xie Yong M, Li Xin, Luo Ji, Shi Xin Q, Hong Xin, Pan Ying H, Ma Xu
Graduate School, Peking Union Medical College, Beijing, China.
Mol Hum Reprod. 2009 Jun;15(6):373-8. doi: 10.1093/molehr/gap028. Epub 2009 Apr 3.
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.
TH2B是一种重要的睾丸组蛋白，在精子发生过程中染色质结构重塑中起关键作用。我们结合高效液相色谱、肽段的酶促Glu-c消化、液相色谱-质谱（LC-MS）和LC-MS/MS分析，对来自精子细胞不同发育阶段的组蛋白TH2B的翻译后修饰（PTM）进行了详细研究。结果显示了精子发生过程中完整组蛋白TH2B的修饰模式。与精母细胞（8.3%）和圆形精子细胞（11.2%）相比，乙酰化的TH2B在精原细胞中最为丰富（28.9%）。鉴定出了TH2B的几种新的PTM。在精原细胞、精母细胞和圆形精子细胞中，T116和K117分别被磷酸化和甲基化修饰，形成了一个新的“磷酸开关”位点。在生精细胞中鉴定出的组蛋白TH2B修饰模式为未来精子发生过程中组蛋白编码和表观遗传调控的研究提供了基础。
