RUI: Defining Structural Features of C. elegans histone H2A Variants Important for Transcriptional Regulation in Different Cell Types

智力优势：不同类型的细胞依赖于不同基因组的表达来实现其发育和功能。改变不同基因组的可接近性是基因调控的一种方式。基因的可接近性可以依赖于将组蛋白H2 A变体掺入到压缩DNA的核小体中。例如，C.线虫表达H2 A变体，包括进化上保守的H2A.Z（HTZ-1）和精子特异性HTAS-1。该项目将测试HTZ-1和HTAS-1的结构差异改变DNA可及性以细胞类型特异性方式调节基因表达的假设。首先，生物物理方法将定义C的结构特性。线虫H2 A蛋白。第二，荧光共振能量转移（FRET）和生物化学方法将确定在稳定性，DNA解包裹，并含有H2 A，HTZ-1和HTAS-1的核小体包装的差异。在遗传突变体中进行的细胞学测定将关联H2 A变体掺入如何影响基因表达。在体外和体内用H2 A交换HTZ-1和HTAS-1的结构域将鉴定这些H2 A变体的结构域，这些结构域对于它们在不同细胞类型中的掺入和转录中的作用是重要的。总之，这些研究将阐明C. elegans组蛋白H2 A变体，其有助于不同细胞类型的特化。更广泛的影响：这项研究将在旧金山弗朗西斯科州立大学进行，这是一个历史上为少数民族服务的公共机构。来自生物学和化学和生物化学课程的本科生和硕士生将由PI和她在SFSU和UCSF的合作者在生物化学，生物物理学和分子遗传学方法的理论和实践中进行培训，以了解DNA组织和转录调控的基本方面。学生将在队列工作，分享资源，想法和专业知识。在PI的监督下，学生将学习批判性地评估科学研究，计划实验，并解释实验数据，成为独立的批判性思考者。拟议工作提供的跨学科培训和指导将为推进SFSU不同学生在科学领域的教育和职业生涯奠定坚实的基础。

Intellectual Merit: Different types of cells rely on the expression of distinct sets of genes for their development and function. Altering the accessibility of different sets of genes is one way genes can be regulated. The accessibility of genes can rely on incorporating histone H2A variants into the nucleosomes that compact DNA. For example, C. elegans express H2A variants that include the evolutionarily conserved H2A.Z (HTZ-1) and the sperm-specific HTAS-1. This project will test the hypothesis that structural differences in HTZ-1 and HTAS-1 alter DNA accessibility to regulate gene expression in a cell-type specific manner. First, biophysical approaches will define structural properties of C. elegans H2A proteins. Second, fluorescence resonance energy transfer (FRET) and biochemical approaches will identify differences in the stability, DNA unwrapping, and packing of nucleosomes containing H2A, HTZ-1, and HTAS-1. Cytological assays conducted in genetic mutants will correlate how H2A variant incorporation affects gene expression. Swapping domains of HTZ-1 and HTAS-1 with H2A in vitro and in vivo will identify structural domains of these H2A variants that are important for their incorporation and roles in transcription in distinct cell types. Overall, these studies will elucidate the structural properties of C. elegans histone H2A variants that contribute to the specification of different cell types. Broader Impacts: The research will be conducted at San Francisco State University, a historically minority-serving public institution. Undergraduate and master's students from the Biology and Chemistry and Biochemistry programs will be trained by the PI and her collaborators at SFSU and UCSF in the theory and practice of biochemical, biophysical, and molecular genetic methodology to understand fundamental aspects of DNA organization and transcriptional regulation. Students will work in cohorts to share resources, ideas, and expertise. Under the supervision of the PI, students will learn to critically evaluate scientific research, plan experiments, and interpret experimental data to become independent critical thinkers. The interdisciplinary training and mentorship provided by the proposed work will provide a strong foundation to advance the education and careers of SFSU's diverse students in the sciences.