NUCLEOSOME POSITIONING

细胞功能的底物，包括转录， 复制、重组和染色体分裂是染色质，而不是 裸露的DNA。这些职能对于发展和健康是必不可少的。 所有的有机体。在染色体组织的初始水平上，DNA是 被压缩成重复排列的核小体。详细的定位 DNA上的核小体对其他阴性或阳性的人来说是必不可少的 基因调控。核小体独特的局部排列是 在已知基因附近的端粒和着丝粒上报道 调节区，以及其他染色体位置；这些特定的 排列被认为是染色体功能所必需的。这个 DNA本身的序列强烈地偏向于核小体的位置 DNA被包裹其中，以及某些DNA序列规则或基序 参与核小体位置的研究已经阐明。这些发现意味着 基因组DNA序列的进化是为了促进它们的功能 通过对其染色质结构的影响。这样做的长期目标是 我们的工作是阐明分子结构之间的关系。 染色体和它们的功能。本项目的目标是 探索基因组DNA序列与核小体的关系 定位和染色体功能。可能关键的事实还没有出现。的确有 需要系统的和定量的分析，以及需要新的 实验工具。我们将通过三种方式实现这些目标。这些 研究建立在我们最近完成的SLEX实验基础上，该实验产生了 大量非天然DNA序列具有极高的亲和力 对于组蛋白八聚体，因此，异常强大的核小体 定位力。(I)我们会采取类似的方法，从 整个酵母基因组这些区域对DNA的作用最大 序列定向的核小体定位。这些序列将是 表征了它们的核小体定位的自由能， 并分析了它们对染色体功能的贡献。(Ii)我们会使用 我们现有的精选定位序列进行操作和研究 体内的核小体定位，我们将使用生物物理方法 以分析DNA的结构和机械性能 负责他们强大的定位能力。(三)我们两个都要 进化和设计方法，以创造下一代Even 更强的定位序列，用于未来的体外和体内研究 活着。

The substrates for cellular functions including transcription, replication, recombination, and chromosome division are chromatin, not naked DNA. These functions are essential for the development and health of all organisms. In the initial level of chromosome organization, DNA is compacted into a repeating array of nucleosomes. The detailed positioning of nucleosomes along DNA can be essential for other negative or positive gene regulation. Distinctive local arrangements of nucleosomes are reported at telomeres and centromeres, in the vicinity of known gene regulatory regions, and at other chromosomal locations; these particular arrangements are believed to be essential for chromosome function. The sequence of the DNA itself strongly biases the position of the nucleosomes into which the DNA is wrapped, and certain DNA sequence rules or motifs involved in nucleosome position have been elucidated. These findings imply that genomic DNA sequences are evolved to facilitate their function through effects on their chromatin structure. The long term aim of this work is to elucidate the relationship between the molecular architecture of chromosomes and their function. The goal of the present project is to explore the relationships between genomic DNA sequence, nucleosome positioning, and chromosome function. May key facts are missing. There is a need for systematic and quantitative analyses, and a need for new experimental tools. We will attack these goals in three ways. These studies build on our recent completion of SLEX experiment which yielded a large number of non-natural DNA sequences have exceptionally high affinity for histone octamer and, consequently, exceptionally strong nucleosome positioning power. (i) We will take similar approaches to select from the entire yeast genome those regions have the greatest power for DNA sequence-directed nucleosome positioning. These sequences will be characterized, their free energy for nucleosome positioning quantified, and their contributions to chromosome function analyzed. (ii) We will use our existing selected positioning sequences to manipulate and study nucleosome positioning in vivo and we will use biophysical approaches in vitro to analyze the DNA structural and mechanical properties that are responsible for their strong positioning power. (iii) We will take both evolutionary and design approaches to create a next generation of even stronger positioning sequences for use in future studies in vitro and in vivo.