Deciphering mechanisms governing functional partitioning of the C. elegans genome

描述(由申请人提供)： 三维基因组组织，其中线性DNA序列被划分为功能不同的区域，对于在每种细胞类型中精确部署遗传信息是必不可少的。最近，越来越多的破坏性疾病和疾病被归因于基因组组织的破坏，但其机制基础仍然鲜为人知，特别是在体内。特别是，结合因子和序列元件的活性如何转化为表观遗传状态和三维基因组结构域来控制基因表达仍然是个谜。线虫提供了一种实验上有利的体内模型来剖析控制基因组结构域正确建立的机制。手头的数据指出了一种新的机制，在这种机制中，序列特异性因子(SNAP190)与RNA聚合酶III复合体的组件共同作用，定义了一个大的基因组结构域，表现出对数千个非编码小RNA的组织特异性、协调调节。在这个提案中，线虫强大的遗传和基因组工具将被用来研究SNAP190、RNA聚合酶III、粘附素和其他候选因子如何与染色质状态(目标1)、染色体环(目标2)和顺式元件(目标3)相互作用，以定义这个独特基因组区域的边界，并在其中调节基因表达。拟议中的实验的成功完成将阐明在体内驱动大规模基因组组织的鲜为人知但至关重要的机制。

DESCRIPTION (provided by applicant): Three-dimensional genome organization, in which the linear DNA sequence is partitioned into functionally distinct domains, is essential for the precise deployment of genetic information in every cell type. An increasing number of devastating disorders and diseases have recently been attributed to disruption of genome organization, yet the mechanistic underpinnings remain poorly understood, especially in vivo. In particular, how the activity of binding factors and sequence elements is translated to epigenetic states and three-dimensional genomic domains to govern gene expression remains mysterious. C. elegans provides an experimentally advantageous in vivo model to dissect the mechanisms governing proper establishment of genomic domains. Data in hand points to a novel mechanism in which a sequence-specific factor (SNAP190) acts with components of the RNA polymerase III complex to define a large genomic domain exhibiting tissue-specific, coordinated regulation of thousands of noncoding small RNAs. In this proposal, the powerful genetic and genomic tools of C. elegans will be employed to investigate how SNAP190, RNA polymerase III, cohesin, and other candidate factors, interact with chromatin states (Aim 1), chromosomal looping (Aim 2), and cis- elements (Aim 3) to define the boundaries of, and regulate gene expression within, this unique genomic domain. The successful completion of the proposed experiments will illuminate the poorly understood yet vitally important mechanisms driving large-scale genome organization in vivo.