GLOBAL ANALYSIS OF HISTONE SUBTYPE COMPOSITION IN C ELEGANS SPERM USING MUDPIT

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 精子是一种特殊的细胞，专门负责将父亲的基因组传递给卵母细胞，而胚胎则由快速分裂的、大部分未分化的细胞组成。与胚胎不同的是，精子已经抛弃了大多数细胞成分，具有高度致密的染色质，并且被认为在很大程度上没有转录活性。因此，精子依赖于替代组蛋白和现有蛋白质的翻译后修饰（PTM）来发育和发挥功能。在其他生物体中，S期组蛋白被组蛋白变体和鱼精蛋白取代，从根本上改变了核小体水平的染色体结构。这两种组织类型之间的染色质差异在很大程度上尚未探索。 探讨C.为了研究线虫精子染色质组成，我们使用多维蛋白质鉴定技术（MudPIT）质谱对精子和胚胎染色质蛋白进行了全面分析。 我们确定不同细胞类型之间染色质组成的新方法已经从胚胎和精子中分别鉴定了1000多种蛋白质，为分析其他组蛋白亚型以及鱼精蛋白直系同源物奠定了基础。此外，我们正在进行的组蛋白PTM的大规模鉴定是研究父系表观遗传标记如何促进胚胎发育的宝贵资源。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Sperm are remarkable cells specialized for the delivery of the paternal genome to the oocyte, while embryos consist of rapidly dividing, largely undifferentiated cells. Unlike embryos, sperm have jettisoned most cellular components, have highly compacted chromatin, and are thought to be largely transcriptionally inactive. Thus, sperm rely on substitution histones and post-translational modification (PTM) of existing proteins for development and function. In other organisms, replacement of S-phase histones by histone variants and protamines fundamentally changes chromosome architecture at the nucleosome level. How chromatin differs between these two tissue types has been largely unexplored. To investigate the nature of C. elegans sperm chromatin composition, we conducted a global analysis of sperm and embryo chromatin proteins using Multidimensional Protein Identification Technology (MudPIT) mass spectroscopy. Our novel approach to determine chromatin composition between disparate cell types has identified over 1000 proteins from embryos and sperm each, laying the groundwork for analysis of other histone subtypes as well as protamine orthologs. In addition, our ongoing large-scale identification of histone PTMs is a valuable resource to examine how paternal epigenetic marks contribute to embryonic development.