RUI: Subcellular Targeting of Protein Kinase C and a Novel Membrane Protein in Polarized Growth of Aspergillus nidulans

RUI：蛋白激酶 C 和新型膜蛋白在构巢曲霉极化生长中的亚细胞靶向

• 批准号: 0742907
• 负责人: Terry Hill
• 金额: $ 47.3万
• 依托单位: Rhodes College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0742907&HistoricalAwards=false
• 关键词: RUI; Subcellular; Targeting; Protein; Kinase

Intellectual Merit The long-term goal of this research is to add to current understanding of the basic mechanisms of growth in the filamentous fungi. Fungi are a large and important group of microorganisms with major impacts on human affairs, through their ability to cause disease in humans and crops and through their usefulness in the production of foods and pharmaceuticals. Our ability to control the activities of both the harmful and the beneficial fungi can be improved through a better understanding of the cellular mechanisms underlying their growth and development. This project will investigate the functions of two proteins, which only recently have been shown to play roles in regulating the growth of filamentous fungi: protein kinase C (PKC), whose activity is essential to the structural integrity of fungal cell walls, and SccA, which is unique to filamentous fungi, and whose function interacts with that of PKC. The specific goals of this research are to determine how PKC and SccA interact with other proteins whose functions in cell wall metabolism are already understood and to elucidate the mechanisms by which PKC and SccA target to sites of cell wall growth. This will be accomplished in part by analyzing how the subcellular distributions of the two proteins are affected when fluorescently labeled (GFP tagged) versions of PKC and SccA are expressed in strains that carry mutations in genes for selected proteins that also function in wall metabolism. Reciprocal experiments will determine how the targeting and function of some of these same wall metabolism proteins are affected by normal and abnormal expression of PKC and SccA. By combining this information with time-lapse observations of the changing distributions of all studied proteins during development of wild type cells, predictive models will be created to guide further research into the interactions between PKC & SccA and other developmentally significant proteins. A second major goal of the research is to analyze how the various structural domains of PKC and SccA contribute to their ability to target correctly to sites of action in the cell and to perform their metabolic functions. This component of the project will involve creating site-mutated or truncated versions of PKC and SccA, and comparing their functions and distribution patterns to those of their normally constructed counterparts.Broader Impact Undergraduate college students will play essential and central roles in all aspects of this research, thus gaining experience in modern molecular biology and becoming more able to make an informed choice about pursuing research careers. In addition to improving the infrastructure of research and education at Rhodes College, the project expands upon a successful existing collaboration between the awardee laboratory and four Historically Black Colleges in the Memphis, Tennessee area. Students from Rust College and Tougaloo College in Mississippi and Lemoyne-Owen College and Lane College in Tennessee will partner with their peers from Rhodes College in team-based research. The project thus strengthens the connections between the participating colleges, while expanding the range of research opportunities available to students from groups traditionally under-represented in the sciences.

智力优势 这项研究的长期目标是增加目前对丝状真菌生长基本机制的理解。 真菌是微生物的一个大而重要的群体，通过它们在人类和作物中引起疾病的能力以及通过它们在食品和药品生产中的用途，对人类事务产生重大影响。 我们控制有害和有益真菌活动的能力可以通过更好地了解它们生长和发育的细胞机制来提高。 该项目将研究两种蛋白质的功能，这两种蛋白质最近才被证明在调节丝状真菌的生长中发挥作用：蛋白激酶C（PKC），其活性对真菌细胞壁的结构完整性至关重要，以及SccA，这是丝状真菌所特有的，其功能与PKC相互作用。 本研究的具体目标是确定PKC和SccA如何与其他蛋白质相互作用，这些蛋白质在细胞壁代谢中的功能已经被理解，并阐明PKC和SccA靶向细胞壁生长位点的机制。这将部分通过分析两种蛋白质的亚细胞分布如何受到影响时，荧光标记（GFP标记）的PKC和SccA的版本在菌株中表达，携带突变的基因选择的蛋白质，也在壁代谢功能。 相互实验将确定这些相同的壁代谢蛋白中的一些的靶向和功能如何受到PKC和Scca A的正常和异常表达的影响。 通过将这些信息与野生型细胞发育过程中所有研究蛋白质分布变化的延时观察相结合，将创建预测模型，以指导进一步研究PKC SccA与其他发育重要蛋白质之间的相互作用。 该研究的第二个主要目标是分析PKC和SccA的各种结构域如何有助于它们正确靶向细胞中的作用位点并执行其代谢功能。 该项目的这一部分将涉及创建PKC和SccA的位点突变或截短版本，并将其功能和分布模式与正常构建的对应物进行比较。 本科生将在这项研究的各个方面发挥重要和核心作用，从而获得现代分子生物学的经验，并更能够对从事研究事业做出明智的选择。 除了改善罗兹学院的研究和教育基础设施外，该项目还扩展了获奖实验室与田纳西州孟菲斯四所历史悠久的黑人学院之间的成功合作。 来自密西西比的Rust学院和Tougaloo学院以及田纳西州的Lemoyne-Owen学院和Lane学院的学生将与来自罗兹学院的同龄人合作进行团队研究。 因此，该项目加强了参与学院之间的联系，同时扩大了传统上在科学领域代表性不足的群体的学生的研究机会。