Analyzing allele specific gene expression and regulation in C. albicans.

分析白色念珠菌的等位基因特异性基因表达和调控。

• 批准号: 7935218
• 负责人: Sadie Lynne Marjani
• 金额: $ 5.22万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-14 至 2011-09-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7935218
• 关键词: Affect; Alleles; Binding; Biological Assay; Biological Models; Cancer Patient; Candida albicans; Chromosomes; DNA Sequence; Disease; Eukaryota; Gene Expression; Gene Expression Regulation; Genomic Instability; Growth; Loss of Heterozygosity; Malignant Neoplasms; Methods; Organism; Pathogenesis; Regulation; Research Design; Role; Tissue-Specific Gene Expression; Transcription Factor 3; Transcriptional Regulation; Work; chromatin immunoprecipitation; fitness; gene replacement; insight; pathogen; transcription factor

DESCRIPTION (provided by applicant): Background: Most eukaryotes are heterozygous, comprised of two distinct alleles. However, little is known about how distinct alleles are differentially expressed and regulated and whether differential regulation can confer fitness to an organism. The proposed studies will provide important information as to the extent of allele specific expression, its mechanisms and function. These topics are highly relevant to understanding mechanisms of cancer. Objective/hypothesis: The natural heterozygosity of C. albicans has allowed it to evolve and adapt to thrive in its mammalian host. However, the degree of differential allelic expression and the mechanism of its regulation are yet to be fully elucidated. I propose to determine the extent of differential allelic expression, and to elucidate if differential binding of specific transcription factors under fungal dimorphic conditions regulates this differential allelic expression. This work is expected to provide insights into the extent and role of heterozygosity of eukaryotes in general. Specific Aims: 1) Globally examine the extent of differential allelic expression in Candida albicans. 2) Determine if differential gene expression is associated with differences in binding of Poll I and transcription factors. 3) Assay the function of differentially regulated alleles. Study Design: I plan to study differential allelic expression and regulation using C. albicans as a model system. I will use 454 DNA sequencing methods to globally analyze allele specific gene expression. ChlPSeq. (chromatin immunoprecipitation and sequencing) will be used to assess how much is due to transcriptional regulation. Finally, I will determine the function of differential allelic expression using gene replacement methods. Relevance: A hallmark of cancer is genome instability. This can be caused by hypomethylation and/or loss of heterozygosity. Moreover, it is common for specific chromosomes and specific chromosomal regions to be preferentially amplified. Thus, basic understanding of the diversity of allele specific expression and how it can be controlled is of high importance to cancer. Moreover, C. albicans is an opportunistic pathogen that affects many diseases, including cancer. Understanding mechanisms that control its growth and switch to dimorphic growth, which is involved in pathogenesis, is of significance for treatment of cancer patients.

描述（由申请人提供）： 背景：大多数真核生物是杂合的，由两个不同的等位基因组成。然而，人们对不同等位基因如何差异表达和调节以及差异调节是否可以赋予生物体适应性知之甚少。拟议的研究将提供有关等位基因特异性表达的程度、其机制和功能的重要信息。这些主题与理解癌症机制高度相关。目的/假设：白色念珠菌的天然杂合性使其能够进化并适应在哺乳动物宿主中繁衍生息。然而，差异等位基因表达的程度及其调控机制尚未完全阐明。我建议确定差异等位基因表达的程度，并阐明真菌二态性条件下特定转录因子的差异结合是否调节这种差异等位基因表达。这项工作预计将提供对真核生物杂合性的程度和作用的见解。具体目标： 1) 全面检查白色念珠菌中差异等位基因表达的程度。 2) 确定差异基因表达是否与 Poll I 和转录因子结合的差异相关。 3) 测定差异调节等位基因的功能。研究设计：我计划使用白色念珠菌作为模型系统来研究差异等位基因表达和调控。我将使用 454 个 DNA 测序方法来全局分析等位基因特异性基因表达。 ChlPSeq。 （染色质免疫沉淀和测序）将用于评估转录调控的程度。最后，我将使用基因替换方法确定差异等位基因表达的功能。 相关性：癌症的一个标志是基因组不稳定。这可能是由低甲基化和/或杂合性丧失引起的。此外，优先扩增特定染色体和特定染色体区域是常见的。因此，对等位基因特异性表达的多样性及其控制方法的基本了解对于癌症非常重要。此外，白色念珠菌是一种机会性病原体，影响许多疾病，包括癌症。了解控制其生长并转变为参与发病机制的二态生长的机制对于癌症患者的治疗具有重要意义。