The goal of this project is to understand how membrane morphology is regulated by the septin cytoskeleton and by lipid composition. To study this question, we use the S. cerevisiae prospore membrane as a model. During sporulation, four new cells are generated within the mother cell as the prospore membrane grows to encapsulate the meiotic nuclei. Ultimately, the prospore membrane will act as the template for the deposition of the spore wall, and its inner leaflet will become the plasma membrane for the new haploid cells. Synthesis of the prospore membrane is essential for yeast to form functional, environmentally resistant spores. Our long-term goal is to understand the regulation of cellular architecture. The proposed research will contribute to our understanding of the molecular mechanisms important for shaping a cellular structure (the prospore membrane) important for the integrity and form of the yeast's meiotic products. The work in this proposal wil provide insights into fundamental aspects of cell biology concerning how cells and cellular structures acquire their shapes, and will also contribute to our understanding of septins, which are filament- forming proteins associated with membranes found in fungi and animals. In the previous funding period, the Spo71 protein was found to be important for determining the size of the prospore membrane and to be required for proper localization of septins along the prospore membrane. Preliminary studies suggest SPO71 is important in the elongation phase of prospore membrane development. The proposed experiments specifically examine the role of septins and septin regulators in prospore membrane development, and also examine how membrane lipid composition is regulated and contributes to prospore membrane morphogenesis. These studies will distinguish models for how these proteins work together to regulate PSM size and shape. As septins are an evolutionarily conserved protein family vital for a variety of cellular processes (including cell division, immune function, and neuronal development) and because the effect of membrane composition on membrane morphology is a fundamental issue in cell biology, the findings from these proposed studies should be of interest to a wide range of biologists.
This proposal investigates how membrane structure is determined by the lipid composition of a membrane and by the proteins that interact with a membrane, a fundamental issue in cell biology. These studies are also of biomedical relevance, as the septin proteins we are examining are membrane-associated filament-forming proteins found in fungi and animals (including humans), and are important for cell division, immune system function, fertility, and nervous system function in mammals. Finally, the membrane we study (the prospore membrane) is critical for the life cycle of many fungal pathogens, and thus these studies may provide insights into fungal-specific mechanisms that can be targeted to combat fungal pathogens that cause human disease.
|Paulissen, Scott M; Huang, Linda S (2016) Efficient Sporulation of Saccharomyces cerevisiae in a 96 Multiwell Format. J Vis Exp :|
|Paulissen, Scott M; Slubowski, Christian J; Roesner, Joseph M et al. (2016) Timely Closure of the Prospore Membrane Requires SPS1 and SPO77 in Saccharomyces cerevisiae. Genetics 203:1203-16|
|Parodi, Emily M; Roesner, Joseph M; Huang, Linda S (2015) SPO73 and SPO71 Function Cooperatively in Prospore Membrane Elongation During Sporulation in Saccharomyces cerevisiae. PLoS One 10:e0143571|
|Slubowski, Christian J; Funk, Alyssa D; Roesner, Joseph M et al. (2015) Plasmids for C-terminal tagging in Saccharomyces cerevisiae that contain improved GFP proteins, Envy and Ivy. Yeast 32:379-87|
|Parodi, Emily M; Baker, Crystal S; Tetzlaff, Cayla et al. (2012) SPO71 mediates prospore membrane size and maturation in Saccharomyces cerevisiae. Eukaryot Cell 11:1191-200|