The interphase microtubule array is a key cellular scaffold that provides structural support and directs organelle trafficking in nearly all eukaryotic cells. Its arrangement is fundamentally important for cell secretion, cell shape, growth, motility, and communication. Although the basic assembly of microtubules is well studied, very little is understood on how the array itself is organized. This is particularly important in multinucleated animal cells where multiple microtubule arrays are present and must maintain spatial separation so as not to interfere with each other. Establishing how these microtubule arrays are organized represents a fundamental challenge in understanding the basic organizational principles of eukaryotic cells. The preliminary data have identified multiple motor and cross-linker proteins that interact with microtubules and when deleted, result in broad alterations in array organization. The research plan is four fold: to understand at the cellular level where these effector proteins operate, to understand at the biochemical level how these effector proteins interact and function, to develop computational models that incorporate these activities and can predict/test understanding in multiple different scenarios, and to provide robust educational experiences to stimulate the next generation of scientists. This project will provide training opportunities for undergraduates recruited from a local teaching college and through Wadsworth's Research Experiences for Undergraduates program, will strengthen research ties with neighboring institutions in the Albany Capitol region, and develop a new working relationship with an expert computational modeler. The investigator will further initiate a mentoring activity for postdocs and new investigators that provides a fresh perspective on career opportunities and pathways in the biological sciences
In detail, the investigator will use combinations of fluorescent reporter fusions and live cell light microscopy to quantitate protein and organelle distributions in multiple mutant backgrounds. This work will provide quantitative values and context for relevant protein interactions in cells. Participants will purify the known effector proteins and develop in vitro assays to determine specific associations and mechanistic detail on how they function. Results obtained from these two objectives will be fed into stochastic agent-based modeling efforts to build simulations that replicate the live cell results, inform whether the investigators need to consider alternate strategies and, importantly, enable prediction on how other microtubule arrays are organized in a broad range of organisms and strategies that are generally understudied in the biosciences. This project will provide training opportunities for undergraduates and mentoring activity for postdocs and new investigators that highlight different career pathways in the biological sciences.
