Analysis of protein dynamics in S. cerevisiae and C. elegans

The temporally and spatially controlled localization of proteins at the plasma is critical for the growth and development of single cellular and multicellular organisms. The balance of secretory and endocytic activities is crucial in determining growth rates, domain establishment and polarity at the plasma membrane. Key regulators of protein and membrane traffic are small GTPases of the ras super family. The activation and inactivation of these small GTPases depend on guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs). The local restriction of these regulatory factors dictates the spatial activation of the GTPases.

In this project, we will determine the functional consequences of activating and inactivating small GTPases of the Arf and Rab families on polarized growth and development. In yeast, we will take advantage of two established light inducible targeting systems to regulate GTPase activity locally through GEFs and GAPs. We will establish GTPase-regulator circuits important for polarized growth in yeast. This knowledge, we will transfer into another highly polarized cell: the C. elegans zygote. We will determine the effect of functionally interfering with the homologous GTPase-regulator circuits on polarity establishment and maintenance from the zygote to the 4-cell stage using RNAi and CRISPR-Cas9 methodology. Finally, we will extend our findings to C. elegans intestinal epithelial cells.

Optogenetics, live cell imaging, molecular biology and biochemical techniques, CRISPR-Cas9, standard yeast and C. elegans techniques.

For more details please visit: https://polarnet-itn.eu/
As part of the project, the successful applicant will visit collaborating labs with in the PolarNet network to learn new techniques.