Project Details
Projekt
Synthetic control of actin nucleation in vivo
Applicant
Professor Dr. Roland Wedlich-Söldner
Subject Area
Cell Biology
Term
from 2013 to 2018
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 239069522
The ever growing number and structural variety of actin nucleation factors poses a huge challenge to structure-function studies. Many factors influencing actin polymerization in cells, such as subcellular protein distribution or presence of biological membranes cannot be easily reproduced in biochemical assays. In addition, a host of actin regulators such as profilin, thymosin, CAP or cofilin influence polymerization kinetics in vivo. However, a cellular system where different nucleators can be characterized under comparable physiological conditions has so far been lacking. We therefore propose to use the budding yeast S. cerevisiae as a minimal physiological system to quantitatively test actin nucleators in vivo. We will first establish a robust assay and controlled cellular environment to monitor actin nucleation at the cortex of budding yeast cells. Using this system we will initially quantitatively characterize the three endogenous yeast nucleators and their regulators. We will then expand our analysis to systematically characterize and compare all types of nucleators from various sources, as well as modified and synthetic actin nucleators. Technically we will combine powerful yeast genetics and molecular biology to precisely manipulate expression and localization of actin nucleators (top down synthetic biology), with live cell Total Internal Reflection Fluorescence Microscopy (TIRFM) and 2D deconvolution to monitor actin nucleation at high temporal and spatial resolution. With our study we aim to provide a reference for the systematic description of actin nucleators.
DFG Programme
Priority Programmes
