RUI: Evolution of Phosphate Starvation Response in Yeast
RUI:酵母中磷酸盐饥饿反应的演变
基本信息
- 批准号:1121714
- 负责人:
- 金额:$ 45.76万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Standard Grant
- 财政年份:2011
- 资助国家:美国
- 起止时间:2011-09-01 至 2015-02-28
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Intellectual merit. This project investigates how the phosphate signal transduction (PHO) pathway changes over evolutionary time in the Ascomycota lineage, with the goal of understanding how evolutionary transitions in this pathway alter growth in different niches. Using the yeasts Candida glabrata and Schizosaccharomyces pombe, these studies focus on the regulation of transcription, elucidation of the ancestral eukaryotic PHO pathway, and how phosphate starvation in all eukaryotes is sensed. Studies with C. glabrata will determine the binding specificity of the transcription factor Pho4. Pho4-regulated promoters in C. glabrata do not contain canonical CACGTG sequences and must utilize different sequences relative to Saccharomyces cerevisiae. The hypothesis that CgPho4 has multiple specificities for PHO promoters will be tested with in vivo (expression from promoter fragments) and in vitro (mobility shift) approaches. This project will capitalize on previous studies demonstrating the neofunctionalization of a C. glabrata acid phosphatase protein, Pmu2. Domains of Pmu2 will be combined with ancestral Pmu1 domains and phosphatase kinetics will be measured. The changes in protein sequence allowing for neofunctionalization will be identified and should be applicable to understanding how new functions evolve more generally. Studies with S. pombe will focus on the identification of Pho7 as a likely transcription factor mediating the phosphate starvation response in S. pombe. The hypothesis that Pho7 binds PHO promoters will be tested utilizing in vivo and in vitro approaches. Finally, the PHO pathway from Chlamydomonas (a green algae) and S. pombe will be genetically reconstituted in S. cerevisiae, allowing for the determination of whether common metabolites in all eukaryotes signal phosphate starvation. Phosphate starvation impacts important cell biological responses and altering the uptake of phosphate for bioremediation or altering the pathogenicity of bacteria and fungi would benefit society. The proposed work will advance our knowledge of transcription factor specificity, the process of neofunctionalization, the possible conservation of metabolic signals of phosphate starvation in all eukaryotes, and a deeper understanding of the evolutionary transitions required in pathways for speciation. Broader impacts. These studies are accessible to relatively new scientists such as undergraduate students and Master's level students. The experiments are parsed into small projects suitable for undergraduates, such as protein purification, enzymatic assays, and plasmid construction. Students will gain valuable experience working with genetics, strain construction, molecular biological techniques, sterile techniques, and biochemistry. Ownership of a task within the framework of a larger project is valuable for students' self-confidence and encourages future self-reliance. This work will support numerous undergraduate and Master's students at Villanova University who will go on to perform scientific research. Students from disadvantaged backgrounds are actively recruited and encouraged to stay in science by positive research experiences and strong mentorship.
智力上的优点。 该项目研究了磷酸盐信号转导(PHO)途径如何在子囊菌门谱系中随着进化时间的推移而变化,目的是了解该途径中的进化转变如何改变不同生态位的生长。 使用酵母光滑念珠菌和裂殖酵母粟酒裂殖酵母,这些研究集中在转录的调节,阐明祖先真核PHO途径,以及如何在所有真核生物的磷酸饥饿感。 研究C。glabrata将决定转录因子Pho4的结合特异性。 C.中Pho 4调节的启动子。glabrata不含有典型的CACGTG序列,并且必须利用相对于酿酒酵母不同的序列。CgPho4对PHO启动子具有多种特异性的假设将用体内(从启动子片段表达)和体外(迁移率变化)方法进行测试。 这个项目将利用以前的研究表明,新功能化的C。光滑型酸性磷酸酶蛋白Pmu 2。 将Pmu2结构域与祖先Pmu1结构域组合,并测量磷酸酶动力学。 允许新功能化的蛋白质序列的变化将被确定,并且应该适用于更普遍地理解新功能是如何进化的。 研究S。pombe将集中于鉴定Pho7作为一个可能的转录因子介导的磷饥饿反应在S.粟酒 Pho7结合PHO启动子的假设将利用体内和体外方法进行测试。 最后,从衣原体(一种绿色藻类)和S.粟酒裂殖酵母将在S.酿酒酵母,允许确定是否在所有真核生物中的共同代谢物的信号磷酸盐饥饿。 磷酸盐饥饿影响重要的细胞生物学反应,改变磷酸盐的吸收用于生物修复或改变细菌和真菌的致病性将有益于社会。 拟议的工作将推进我们的转录因子特异性的知识,新功能化的过程中,在所有真核生物的磷酸盐饥饿的代谢信号的可能的保护,并更深入地了解物种形成的途径所需的进化转变。更广泛的影响。这些研究是相对较新的科学家,如本科生和硕士水平的学生。 这些实验被分解成适合本科生的小项目,如蛋白质纯化,酶测定和质粒构建。学生将获得与遗传学,菌株建设,分子生物学技术,无菌技术和生物化学工作的宝贵经验。 在一个更大的项目框架内拥有一项任务对学生的自信心很有价值,并鼓励未来的自力更生。 这项工作将支持许多本科生和硕士生在维拉诺瓦大学谁将继续进行科学研究。 来自弱势背景的学生被积极招募,并鼓励他们通过积极的研究经验和强大的导师留在科学领域。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Dennis Wykoff其他文献
Dennis Wykoff的其他文献
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{{ truncateString('Dennis Wykoff', 18)}}的其他基金
RUI: Characterization of thiamine signal transduction pathway in Candida glabrata and other Ascomycetes
RUI:光滑念珠菌和其他子囊菌中硫胺素信号转导途径的表征
- 批准号:
1921632 - 财政年份:2019
- 资助金额:
$ 45.76万 - 项目类别:
Standard Grant
RUI: Evolution of Signal Transduction Pathways in Yeast
RUI:酵母信号转导途径的进化
- 批准号:
1412582 - 财政年份:2014
- 资助金额:
$ 45.76万 - 项目类别:
Standard Grant
RUI: Evolution of the Phosphate Starvation Response in Yeast
RUI:酵母中磷酸盐饥饿反应的演变
- 批准号:
0747799 - 财政年份:2008
- 资助金额:
$ 45.76万 - 项目类别:
Continuing Grant
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