Control of Planar cell polarity and spindle positioning in C. elegans epithelial Seam Cells


Control of Planar cell polarity and spindle positioning in C. elegans epithelial Seam Cells

Supervising PI

Sander van den Heuvel (website )

Project Description

Objectives

Identification of regulatory mechanisms of planar cell polarity and its connection to cell division control.

Methodology

We will use CRISPR/Cas9-mediated genome engineering to generate floxed alleles of C. elegans Wnt/β-catenin asymmetry pathway components, cell junction components, and potential PCP genes. Inducible-tissue specific Cre expression will be used to accomplish gene inactivation in a subset of seam cells. We will examine the orientation of cell polarity and spindle positioning, both in the knock out cells and their neighbours, using a variety of fluorescent marker lines that have been created in our group. Moreover, we will determine whether these signalling and adhesive interactions between neighbouring cells control the timing of cell division and “stemness” cell fate of epithelial seam cells. As a promising novel strategy, we have recently pioneered methods for optogenetic control (light-inducible) protein dimerization in C. elegans. We aim to use this method for spatiotemporal control of protein localization in seam cells, followed by quantitative measurements of spindle positioning and cell division timing.

Collaborators

 Key publications

  1. Ruijtenberg S, van den Heuvel S. G1/S Inhibitors and the SWI/SNF Complex Control Cell-Cycle Exit during Muscle Differentiation. Cell. 2015 Jul 16;162(2):300–13.
  2. The I, Ruijtenberg S, Bouchet BP, Cristobal A, Prinsen MBW, van Mourik T, et al. Rb and FZR1/Cdh1 determine CDK4/6-cyclin D requirement in C. elegans and human cancer cells. Nat Commun. 2015;6:5906.
  3. Wildwater M, Sander N, de Vreede G, van den Heuvel S. Cell shape and Wnt signaling redundantly control the division axis of C. elegans epithelial stem cells. Development. 2011 Oct;138(20):4375–85.
  4. Galli M, Muñoz J, Portegijs V, Boxem M, Grill SW, Heck AJR, et al. aPKC phosphorylates NuMA-related LIN-5 to position the mitotic spindle during asymmetric division. Nat Cell Biol. 2011 Sep;13(9):1132–8.