Rap 1 binding and transcriptional regulation in meiosis

描述（由申请人提供）：Rap 1 p是酿酒酵母的一种必需DNA结合蛋白，在沉默大片段染色质以及激活和抑制数百个基因中发挥作用。在快速生长的酵母细胞中，许多由Rap 1 p调控的基因是基因组中转录最高的;然而，在其他生长条件下，包括减数分裂，许多相同的基因被抑制。了解单个转录因子如何根据细胞类型、发育阶段或生长条件介导不同的转录反应是一个基本的生物学问题。本提案中描述的实验旨在确定与植物生长细胞相比，允许Rap 1 p在减数分裂期间引起不同转录结果的因素。首先，Rap 1 p的分布及其对营养生长和整个减数分裂中的表达模式的影响将使用ChIP芯片和表达微阵列实验进行映射。然后，我们将比较每种条件下的分布模式，并将结合模式与基因表达相关联。最后，使用敏感的报告分析系统，我们将进行筛选，以确定和表征辅因子，竞争转录因子和蛋白质修饰剂，需要在减数分裂过程中调节Rap 1 p的活性。

DESCRIPTION (provided by applicant): Rap1p, an essential DNA binding protein of Saccharomyces cerevisiae plays roles in silencing large stretches of Chromatin and in the activation and repression of hundreds of genes. In rapidly growing yeast cells, many of the genes regulated by Rap1p are among the most highly transcribed in the genome; however, during other growth conditions, including meiosis, many of the same genes are repressed. Understanding how a single transcription factor can mediate different transcriptional responses based on cell type, stage of development or growth conditions is a fundamental biological problem. The experiments described in this proposal are designed to identify factors that allow Rap1p to evoke different transcriptional outcomes during meiosis as compared to vegetatively growing cells. First, the distribution of Rap1 p and its affects on patterns of expression in vegetative growth and throughout meiosis will be mapped using ChlP-chip and expression microarray experiments. We will then compare the patterns of distribution under each condition and correlate binding patterns with gene expression. Finally, using sensitive reporter assay systems, we will perform screens to identify and characterize cofactors, competing transcription factors and protein modifiers that are required for modulating the activity of Rap1p during meiosis.