睾丸特有的核小体变体及其调控因子的结构与动力学研究

During spermatogenesis, postmeiotic male germ cells undergo extensive chromatin remodeling. This process involves large-scale incorporation of testis specific histone variants, creating less stable nucleosomes; genome-wide histone hyperacetylation; and the testis specific chromatin remodeling protein factors function. That causes chromatin more open in certain areas. We will study the testis-specific histone variants, histone chaperone and chromatin remodeling protein factor by structural biology methods to answer following questions:.1..The testis-specific histone variants substituted in the nucleosome, how to influence nucleosome structure and stability? What are its dynamic properties?.2. Histone chaperone related with spermatogenesis, how they interact with the nucleosome. How they perform their function?.3. The structure of testis-specific chromatin remodeling protein factor that related with histone acetylaion, how they interact with the nucleosome. How they perform their function?.Our research will promote the structural biology research of epigenetic regulation from studying interaction between protein and peptide to the level interaction between protein and nucleosome. It also will promote the NMR studying from small proteins (MW <20 kDa ) to supermolecules. During the investigation, we will also concerning methodology development.. In the structural biology investigation for epigenetic regulation, our laboratory already has solid background. We have published series papers in international Journal such as Genes & Developmen, JBC, J. strut.Bio., et al. Papers were cited by many important journal that publish review articles.

我们将研究精子形成过程中，睾丸中特有的组蛋白变体组装的核小体；组蛋白的分子伴侣；染色质重塑蛋白质因子的结构，动力学。回答以下科学问题：.1、睾丸中特有的组蛋白变体组装进核小体，如何影响核小体的结构及稳定性？其动力学特征是什么？2、与精子形成过程相关的组蛋白分子伴侣的结构，它们与核小体如何相互作用，行使功能？3、睾丸中特有的与组蛋白乙酰化相关的染色质重塑蛋白因子的结构，它们与核小体如何相互作用，行使功能？通过研究，力图将表观遗传调控的结构生物学研究由研究蛋白质-多肽相互作用，推向研究蛋白质与核小体作用；将核磁共振研究由小蛋白质推向超大分子；方法技术有所创新。我们实验室在表观调控的结构生物学研究方面具有相当基础。在Genes & Developmen, JBC, J. strut.Bio. 等学术期刊发表一批论文，文章被许多重要综述期刊引用。

精子形成过程中，染色质发生最为剧烈的重塑，染色质的高级结构要有序地受到外部因子如染色质重塑复合物，DNA及组蛋白修饰酶，组蛋白分子伴侣等外部因子的调控。在上述过程中核小体也会组装与去组装，同时伴随不同组蛋白变体的交换。本项目立足于研究精子形成过程中，睾丸中特有的组蛋白变体组装的核小体、组蛋白的分子伴侣、染色质重塑因子的结构和动力学。回答以下科学问题：1、睾丸中特有的组蛋白变体组装进核小体，如何影响核小体的结构及稳定性？其动力学特征是什么？2、与精子形成过程相关的组蛋白分子伴侣的结构，它们与核小体如何相互作用，行使功能？3、睾丸中特有的与组蛋白乙酰化相关的染色质重塑蛋白因子的结构，它们与核小体如何相互作用，行使功能？按照计划已成功组装野生型全氘代重组爪蟾核小体，以及蛋白变体H2A.Bbd的核小体，获得了高纯度的、可开展结构生物学研究的毫克量级的组蛋白变体H2A.Bbd的核小体，开展了睾丸特有的蛋白质Trim66与组蛋白H3K56ac的相互识别，及其功能研究；研究了裂殖酵母减数分裂与有丝分裂转化的关键蛋白质的结构与功能研究。还开展了其它一些与表观遗传调控相关的蛋白质复合物的结构与功能研究。在Nature communication, Nucleic Acids research, Cell research等期刊发表SCI论文27篇，其中中科院分区I区论文占37%，I/II区论文合计占74%。通过研究，将表观遗传调控的结构生物学研究由研究蛋白质-多肽相互作用，推向研究蛋白质与核小体作用，将核磁共振研究由小蛋白质推向超大分子，并在方法技术上有所创新。