Genetic interactions; Image-based screening; Single-cell transcriptomics; Membrane-less compartmentalization; Phase separation; Kinases
PopovicDoris, NijenhuisWilco, KapiteinLukas, PelkmansLucas (2020), Co-translational targeting of transcripts to endosomes, 1.
GalloRaffaella, RaiArpan, PelkmansLucas (2020), DYRK3-Controlled Phase Separation Organizes the Early Secretory Pathway, 1.
D Berchtold, N Battich, L Pelkmans (2018), A Systems-Level Study Reveals Regulators of Membrane-less Organelles in Human Cells., in Molecular cell
AK Rai, JX Chen, M Selbach, L Pelkmans (2018), Kinase-controlled phase transition of membraneless organelles in mitosis., in Nature
de Groot R, J Lüthi, H Lindsay, H Lindsay, R Holtackers, L Pelkmans (2018), Large-scale image-based profiling of single-cell phenotypes in arrayed CRISPR-Cas9 gene perturbation screens., in Molecular systems biology
D Popovic, B Koch, M Kueblbeck, J Ellenberg, L Pelkmans (2018), Multivariate Control of Transcript to Protein Variability in Single Mammalian Cells., in Cell systems
This proposal will study a generalizable molecular regulatory principle of a fundamental but poorly understood process occurring in eukaryotic cells, namely the membrane-less compartmentalization of cyto- and nucleoplasm through liquid phase transitions. It will build on our recent discovery of a cyclic partitioning mechanism displayed by the dual-specificity kinase DYRK3, and extend it to multiple protein kinases. We propose that this cycle allows liquid-liquid unmixing to be regulated and coupled to other cellular processes, comparable to the role of the RabGTPase cycle in the regulation of membrane-bound compartmentalization. An iterative bioinformatics and systematic imaging approach will be applied to identify predictive signatures for liquid-liquid unmixing across all protein kinases. This is followed by molecular cell biology and biochemistry experiments to map the domains and phosphorylation sites involved in the cyclic partitioning mechanism of several representative kinases. Using image-based assays for different types of liquid-liquid unmixed compartments, an innovative gene perturbation screening approach will then be applied to map regulatory genetic interactions underlying liquid-liquid unmixing. Finally, a functional role of kinase-regulated liquid-liquid unmixing in the mesoscale organization of the transcriptome and gene expression will be studied using image-based transcriptomics.