多组学方法揭示组蛋白 H3.1 在精原干细胞体外分化过程中的表观遗传调控

Multi-Omics Approaches for Revealing the Epigenetic Regulation of Histone H3.1 during Spermatogonial Stem Cell Differentiation In Vitro.

机构信息

State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 210029, China.

State Key Laboratory of Reproductive Medicine, Women's Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing Medical University, Nanjing 210029, China.

出版信息

Int J Mol Sci. 2023 Feb 7;24(4):3314. doi: 10.3390/ijms24043314.

DOI:10.3390/ijms24043314

PMID:36834727

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9958608/

Abstract

Epigenetic regulation, particularly post-translational modifications (PTMs) of histones, participates in spermatogonial stem cell (SSCs) differentiation. However, there is a lack of systemic studies of histone PTM regulation during the differentiation of SSCs due to its low number in vivo. Herein, we quantified dynamic changes of 46 different PTMs on histone H3.1 by targeted quantitative proteomics using mass spectrometry during SSCs differentiation in vitro, in combination with our RNA-seq data. We identified seven histone H3.1 modifications to be differentially regulated. In addition, we selected H3K9me2 and H3S10ph for subsequent biotinylated peptide pull-down experiments and identified 38 H3K9me2-binding proteins and 42 H3S10ph-binding proteins, which contain several transcription factors, such as GTF2E2 and SUPT5H, which appear to be crucial for epigenetic regulation of SSC differentiation.

摘要

表观遗传调控，特别是组蛋白的翻译后修饰（PTMs），参与精原干细胞（SSCs）的分化。然而，由于体内 SSCs 的数量较少，对于 SSCs 分化过程中组蛋白 PTM 调控的系统性研究还很缺乏。在这里，我们通过使用质谱的靶向定量蛋白质组学，对体外 SSCs 分化过程中组蛋白 H3.1 上的 46 种不同 PTM 进行了定量，结合我们的 RNA-seq 数据。我们鉴定到 7 种差异调控的组蛋白 H3.1 修饰。此外，我们选择 H3K9me2 和 H3S10ph 进行后续的生物素化肽下拉实验，鉴定到 38 种 H3K9me2 结合蛋白和 42 种 H3S10ph 结合蛋白，其中包含几个转录因子，如 GTF2E2 和 SUPT5H，它们似乎对 SSC 分化的表观遗传调控至关重要。