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.
