RUI: Evolution of the Phosphate Starvation Response in Yeast

学术成就本项目研究子囊菌门真菌谱系中的磷酸盐信号转导（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是保守的，这为进行详细的比较基因组分析提供了难得的机会。本研究的两个目的是：（1）确定光滑假丝酵母的磷饥饿反应和PHO途径;（2）确定粟酒裂殖酵母的PHO途径。 C. glabrata是与S.酿酒酵母和Sz.粟酒裂殖酵母是一种远亲模式生物。 比较S。cerevisiae和C. glabrata PHO通路的研究允许分析信号转导通路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。 将表征PHO途径的上游信号传导组分和下游转录输出。 因为Sz.粟酒裂殖酵母与S.酿酒酵母，但产生了类似的输出（磷酸盐饥饿基因的转录上调），这项研究将提供背景，在一个类似的生态位不同的生物体如何进化出磷酸盐饥饿反应，并采取了类似的基因表达程序与不同的信号通路。八个突变体的磷酸盐调节基因PHO 1的表达异常的特点和磷酸盐饥饿反应所需的基因将被克隆。 将使用DNA微阵列和rt-qPCR测量这些突变体（和野生型）的转录应答。 这些研究适合相对较新的科学家，例如本科生和硕士生。这些系统的比较基因组分析将有助于解释在不同生态位中定制生长途径所需的进化步骤。 在这个项目中使用的大多数技术都是初级科学家可以使用的。 许多实验利用基本的遗传和分子生物学技术，是为本科生和硕士生设计的独立项目。 独立的项目是学生的理想选择，因为它们鼓励学生对整个项目的一个方面的所有权。这项工作将为维拉诺瓦大学的众多本科生和硕士生创造资源，以获得未来生物医学或科学研究科学方法的实践经验。学生也将被强烈鼓励参加科学会议，与其他科学家互动。最后，来自弱势背景和代表性不足的少数群体的学生将被积极招募，并鼓励他们通过积极的本科/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.