RUI: Evolution of the Phosphate Starvation Response in Yeast

智力价值这个项目研究了子囊菌门真菌谱系中的磷酸信号转导(Pho)途径。子囊菌从数亿年前的一个共同的真菌祖先分化而来，酿酒酵母的pho途径的许多方面在谱系中都是保守的，这为进行详细的比较基因组分析提供了一个难得的机会。本项目的两个目标是(1)确定光滑假丝酵母中的磷酸饥饿反应和pho途径；(2)确定裂殖酵母中的pho途径。光肩星天牛是一种模式种，与酿酒酵母和Sz.庞贝是一种远亲关系较远的模式生物。通过比较酿酒酵母和光皮葡萄球菌的途径，可以分析信号转导途径如何改变密切相关物种的转录输出，以及这些改变如何影响生态位特异性。将描述pho途径的上游信号成分和下游转录输出。因为Sz.Pombe与酿酒酵母的pho信号通路没有同源成分，但产生了相似的输出(磷酸盐饥饿基因的转录上调)，这项研究将为相似生态位中的不同生物如何进化出磷酸盐饥饿反应提供背景，并采用不同信号途径的相似基因表达程序。将对8个磷酸盐调节基因PHO1表达异常的突变体进行鉴定，并克隆磷酸盐饥饿反应所需的基因。这些突变体(和野生型)的转录反应将用DNA微阵列和RT-qPCR来测量。这些研究对相对较新的科学家来说是可以获得的，比如本科生和硕士研究生。这些系统的比较基因组分析将有助于解释在不同的生态环境中为生长定制路径所需的进化步骤。广泛影响这个项目中使用的大多数技术对初学者来说都是可用的。许多实验利用基本的遗传和分子生物学技术，是为本科生和硕士研究生设计的独立项目。自给自足的项目对学生来说是理想的，因为它们鼓励学生对整个项目的一个方面拥有所有权。这项工作将为维拉诺瓦大学的众多本科生和硕士学生创造资源，为未来的生物医学或科学研究获得科学方法的实践经验。还将强烈鼓励学生参加科学会议，与其他科学家互动。最后，将积极招募来自弱势背景和代表性不足的少数群体的学生，并鼓励他们通过积极的本科生/硕士经历留在科学领域。

Intellectual MeritThis project investigates the phosphate signal transduction (PHO) pathway in the Ascomycota fungal lineage. Ascomycetes diverged from a common fungal ancestor hundreds of millions of years ago and many aspects of the PHO pathway from Saccharomyces cerevisiae are conserved in the lineage, providing a rare opportunity to perform detailed comparative genomic analyses. The two aims of this project are (1) to define the phosphate starvation response and PHO pathway in Candida glabrata and (2) to define the PHO pathway in Schizosaccharomyces pombe. C. glabrata is a model species closely related to S. cerevisiae and Sz. pombe is a distantly related model organism. Comparing the S. cerevisiae and C. glabrata PHO pathways allows for an analysis of how a signal transduction pathway alters transcriptional output in closely related species and how these alterations impact niche specificity. The upstream signaling components and downstream transcriptional output of the PHO pathway will be characterized. Because Sz. pombe does not share orthologous components with the PHO signaling pathway of S. cerevisiae, but generates a similar output (transcriptional up-regulation of phosphate starvation genes), this research will provide the context for how a divergent organism in a similar niche has evolved a phosphate-starvation response, and adopted a similar gene expression program with a different signaling pathway. Eight mutants aberrant in their expression of the phosphate-regulated gene PHO1 will be characterized and the genes required for the phosphate starvation response will be cloned. The transcriptional response of these mutants (and wild-type) will be measured with DNA microarrays and rt-qPCR. These studies are accessible to relatively new scientists such as undergraduate students and Master's level students. These systematic comparative genomic analyses will help to explain the evolutionary steps required to tailor a pathway for growth in different niches.Broader Impact Most of the techniques utilized in this project are accessible to beginning scientists. Many experiments utilize basic genetic and molecular biology techniques and are self-contained projects designed for undergraduate and Master's level students. The self-contained projects are ideal for students in that they encourage student ownership of an aspect of the total project. This work will create the resources for numerous undergraduates and Master's students at Villanova University to gain practical experience with the scientific method for a future in biomedical or scientific research. Students will also be strongly encouraged to attend scientific meetings to interact with other scientists. Finally, students from disadvantaged backgrounds and underrepresented minority groups will be actively recruited and encouraged to stay in scientific fields through a positive undergraduate/Masters experience.