in vivo tools for functional analysis of microtubule network architecture

Ashleigh Fritz

Understanding the architecture of cytoskeletal networks underlying different epithelial morphologies is critical to understanding the development of tissues and organs.  To expand the molecular toolkit available to test microtubule (MT) function in vivo, we are analyzing the effect of expressing a MT-severing bacterial toxin on MT architecture in Drosophila epithelial cells. Expression of the bacterial gene disrupts the normal cell morphology, leading to more rounded cells and enhanced cell death.  Presently, we are creating a line of flies to co-express the bacterial toxin with p35 to suppress cell death in the MT-deficient cells.  We will also create novel fusion constructs to subcellularly localize the MT-severing activity.  Finally, in collaboration with Jen Gerton, we are attempting to use the Saccharomyces cerevisiae to screen for functional variants of the bacterial protein.