Biochemistry and Molecular Cell Biology

Note: this project takes place on Health Sciences Campus in Maywood, IL

The cytoskeleton is a complex and dynamic network of proteins that primarily function to give cells their characteristic shape and enables them to form complex multicellular structures. It plays a role in everything from cell division and replication to more subtle signaling. The eukaryotic cytoskeleton is largely thought to be comprised of three main components, actin filaments, intermediate filaments and microtubules. More recently it has been argued that we should include a fourth, less well studied filamentous protein, septins, as a component of the cytoskeleton. Septins are the primary focus of my research.

While it is known that septin and actin filaments colocalize in eukaryotic cells, it is unknown whether there is direct overlap between the two, especially at the cortex and in actin fibers around the nucleus. During this summer undergraduate project, I would like my research assistant to look at whether septins overlap with actin filaments using super-resolution microscopy and whether the extent of overlap changes at regions of active RhoA. I would like to test this in three different cell lines – fibroblasts with endogenously tagged Sept2 (control), cells that have reduced expression of septins and cells that overexpress septins. My research assistant will receive training in cell culture, fixing and staining cells to identify different cytoskeletal components and observe the extent of their overlap, under the Airyscan 880 microscope, local and global activation of RhoA followed by fixing and staining the activated cells to observe the extent of overlap between septin and actin filaments.