Regulation of cilia dynamics during the cell cycle Contact Dr. Peter van der Sluijs, Department of Cell Biology, UMC Utrecht, E/T: [email protected]/088-7557580 Introduction Cilia are microtubule-based structures that protrude from the cell surface, and function as sensors for environmental cues that regulate cellular differentiation or division. In metazoans, ciliary signaling is important both during development and in the homeostasis controls of adult tissues. Cilia are dynamically regulated during cell cycle progression. They are present on interphase cells and usually resorbed before mitotic entry, to re-appear post-cytokinesis. It is not clear how this happens. We recently discovered a possible mechanism that involves regulation of polarized protein transport to the cilium. Outline of experimental plan The aim of this project is to investigate regulation of protein transport to the cilium during the cell cycle with transfected GFP-tagged reporters using live cell imaging and single cell biochemistry. Methods - Analysis of living cells with (dual label) fluorescence microscopy - Lentiviral transduction - Fluorescence Activated Cell Sorting - Knock down in tissue culture cells with RNAi - Recombinant DNA methods to prepare expression constructs - Protein methods like SDS-PAGE, Western blot, immunoprecipitation Selected papers from the group - Hoogenraad CC, Popa I, Futai K, Sanchez-Martinez E, Wulf PS, van Vlijmen T, Dortland BR, Oorschot V, Govers R, Monti M, Heck AJR, Sheng M, Klumperman J, Rehmann H, Jaarsma D, Kapitein LC, and van der Sluijs P. 2010. Neuron specific Rab4 effector GRASP-1 coordinates membrane specialization and maturation of recycling endosomes. PLoS Biol. 8: e1000283. - van Vlijmen* T, Rojas* R, Mardones G, Mohammed S, Heck AJR, Raposo G, van der Sluijs P and Bonifacino JS. 2008. Regulation of retromer recruitment to endosomes by sequential action of rab5 and rab7. (* denotes equal contributions). J. Cell Biol.183: 513-526.