Collaborative Research - Role of Pulsatile Ca^2+ in Controlling Polarized Hyphal Tip Growth

智力上的优点。所有生物体都有识别和适应环境的复杂方法，以便它们能够生存或避免恶劣条件，开发新的食物来源，甚至引发疾病。在DNA测序、解码和修饰方法方面的几项最新进展，以及在统一方法中使用高倍显微镜的技术改进，现在可以在活细胞内监测目标基因变化的细微异常影响。这项研究使用真菌作为模型系统，以确定细胞如何能够感知其微观环境的变化，并招募高度专业化的蛋白质，这些蛋白质在随后的一系列事件中与钙离子结合，从而改变细胞的行为。遗传方法将被用来全面破坏真菌模式生物中所有已知的关键钙相互作用蛋白，并附加特殊标签，允许监测钙的变化和/或多个钙相互作用和相关蛋白的运动，以便在活细胞内进行跟踪。这种方法将导致全面了解哪些目标蛋白允许细胞以快速和定向的方式生长。这个项目的科学影响不仅仅是更好地了解模式真菌禾谷镰刀菌中钙信号的生物学，它会导致小麦的一种严重疾病。它将创建一个新的工具箱，用于研究钙信号的进化，钙信号是一种高度保守的古老分子语言，几乎在所有生物体中都使用，将外部环境信号转换为必要的细胞变化。该项目汇集了一群具有互补背景的科学家，包括生物化学、细胞生物学、细胞学、进化生物学、真菌生物学、信息学、数学和遗传学。该项目广泛的跨学科范围将为本科生、研究生和博士后学生实践基于团队的综合问题解决提供理想的环境。面对迅速增长的生物学问题的复杂性，对这种教育的需求继续增加。由该项目支持的一名博士后将在不久的将来通过既定的指导计划开发自己的项目。通过为期一周的实践讲习班和全面的公开网站，其他人将获得所有资源，并接受实施所开发工具的培训。这里开发的钙信号发现和显微技术将在研究生水平的课程和基于网络的工具中介绍。本科生(包括少数族裔学生)将通过NSF、REU和EPSCoR项目以及宾夕法尼亚州立大学和特拉华大学现有的其他学分和外展项目参与这一项目。这些努力将帮助参与的学生获得广阔的视角和新技术，以便在学术和工业环境中推广。这个真菌模型将揭示动植物中其他类似生长形式(即神经元、发芽和分裂酵母、上皮细胞、根毛、花粉管)中钙信号通路的共同组成部分，并揭示与它们的进化适应和功能多样性有关的独特属性。真菌钙信号的独特方面可能被用来控制疾病。

Intellectual merit. All organisms have sophisticated ways for recognizing and adapting to their environment so they can survive or avoid harsh conditions, exploit new food sources or even cause disease. Several recent advances in methods for sequencing, decoding and modifying DNA and technological improvements in powerful microscopes when used in a unified approach now enable subtle abnormal effects of targeted genetic changes to be monitored inside living cells. This research uses fungi as a model system for determining how cells are able to sense changes in their microscopic environment and recruit highly specialized proteins that bind to calcium ions in a subsequent cascade of events that change the cells behavior. Genetic methods will be used to comprehensively disrupt all known key calcium interacting proteins in a fungal model organism and attach special tags that allow monitoring of calcium changes and/or movement of multiple calcium interacting and related proteins to be tracked inside of living cells. This approach will result in a comprehensive understanding of which targeted proteins allow cells to grow in a rapid and directed fashion. The scientific impacts of this project go beyond better understanding the biology of calcium signaling in the model fungus Fusarium graminearum which causes a serious disease in wheat. It will create a novel toolbox for studying the evolution of calcium signaling, a highly conserved ancient molecular language used by in virtually all organisms for converting external environmental signals into needed cellular changes. The project brings together a group of scientists with complementary backgrounds including; biochemistry, cell biology, cytology, evolutionary biology, fungal biology, informatics, math, and genetics.Broader impacts. The broad and interdisciplinary scope of this project will provide an ideal environment for practicing team-based integrative problem solving with undergraduate, graduate, and postdoctoral students. Faced with the rapidly growing complexity of biological questions, the need for such education continues to increase. A postdoc supported by this project will be mentored to develop her own program in the near future through established mentoring programs. Through hands-on week long workshops and comprehensive publicly available websites, others will have access to all resources and trained to implement the developed tools. The calcium signaling discoveries and microscopy techniques developed here will be introduced in graduate level courses and web-based tools. Undergraduate students (including minority students) will be involved in this project through NSF REU and EPSCoR programs as well as other existing credit and outreach programs at both Penn State and University of Delaware. These efforts will help the participating students gain broad perspectives and new technologies to carry forth into academic and industrial settings. This fungus model will reveal shared components of calcium signaling pathways in other similar growth forms (i.e. neurons, budding and fission yeast, epithelial cells, root hairs, pollen tubes) in animals and plants, and also expose unique attributes potentially relating to their evolutionary adaptation and functional diversity. Unique aspects of fungal Ca2+ signaling may be exploited for controlling disease.