Mass spectrometry analysis of dynamic post-translational modifications of TH2B during spermatogenesis.
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.
Graduate School, Peking Union Medical College, Beijing, China.
Shuang
S
Lu
Yong M
YM
Xie
Xin
X
Li
Ji
J
Luo
Xin Q
XQ
Shi
Xin
X
Hong
Ying H
YH
Pan
Xu
X
Ma
D000107:Acetylation; D000818:Animals; D002853:Chromatography, Liquid; D006657:Histones / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008745:Methylation; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley; D013087:Spermatids / Q000378:metabolism; D013090:Spermatocytes / Q000378:metabolism; D013091:Spermatogenesis / Q000502:physiology*; D053719:Tandem Mass Spectrometry
D006657:Histones; C081018:histone H2B type 1-A