Clathrin-mediated endocytosis is an important process involved in uptake of molecules into cells, signalling or membrane homeostasis. At the plasma membrane the clathrin pit assembly and cargo selection is assisted by adaptor proteins, among them the AP2 adaptor protein complex. In mammalian cells AP2 activity is regulated by AAK1 kinase phosphorylation. We utilized the nematode Caenorhabditis elegans to study the developmental role of AAK1 ortholog SEL-5 in this invertebrate model. SEL-5 activity is dispensable for general regulation of endocytosis in C. elegans, however loss of SEL-5 activity leads to a defect in migration of neuronal precursor cells and also to a defect in cell growth of a particular cell. Interestingly, defect in cell migration can also be observed in cultured mammalian cells after loss of AAK1 expression. AAK1 and SEL-5 thus seem to act in both similar and disparate mechanisms. The aim of this project is to dissect the mechanism of SEL-5 function in neuronal migration and in cell growth and compare the AAK1/SEL-5 mechanism of action between mammalian and C. elegans models. The project will combine advanced C. elegans genetic techniques including targeted CRISPR/Cas9 mutagenesis and in vivo protein tagging with biochemical and microscopic analyses in cultured mammalian cells.
Five relevant publications of the research group:
Doubravská, L., Dostál, V., Knop, F., Libusová, L., Macůrková, M. (2020) Human myotubularin-related protein 9 regulates ER-to-Golgi trafficking and modulates WNT3A secretion. Exp Cell Res. 386(1):111709.
Lorenowicz, MJ., Macůrková, M., Harterink, M., Middelkoop, TC., de Groot, R., Betist, MC., Korswagen, HC. (2014) Inhibition of late endosomal maturation restores Wnt secretion in Caenorhabditis elegans vps-29 retromer mutants. Cell Signal. 26(1):19-31.
de Groot, RE, Farin, HF, Macůrková, M, van Es, JH, Clevers, HC, Korswagen, HC. (2013) Retromer dependent recycling of the wnt secretion factor wls is dispensable for stem cell maintenance in the mammalian intestinal epithelium. PLoS One. 8(10):e76971.
Harterink, M., Port, F., Lorenowicz, MJ., McGough, IJ., Šilhánková, M., Betist, MC., van Weering, JR., van Heesbeen, RG., Middelkoop, TC., Basler, K., Cullen, PJ., Korswagen, HC. (2011) A SNX3-dependent retromer pathway mediates retrograde transport of the Wnt sorting receptor Wntless and is required for Wnt secretion. Nat Cell Biol. 13(8):914-23.
Šilhánková, M., Port, F., Harterink, M., Basler, K., Korswagen, HC. (2010) Wnt signalling requires MTM-6 and MTM-9 myotubularin lipid-phosphatase function in Wnt-poducing cells. EMBO J. 29(24):4094-105.