Nuclear immunophilin FKBP39 from Drosophila melanogaster drives spontaneous liquid-liquid phase separation
| Autor: | Krzysztof Wycisk, Magda Drewniak-Świtalska, Łukasz Berlicki, Marek Orłowski, Aneta Tarczewska, Andrzej Ożyhar, Jurek Dobrucki, Agnieszka Waligórska |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Nucleoplasmin
Recombinant Fusion Proteins FKBP39 02 engineering and technology Sodium Chloride Arginine Biochemistry Homology (biology) Mass Spectrometry Tacrolimus Binding Proteins 03 medical and health sciences Protein Aggregates Structural Biology Chlorocebus aethiops Animals Drosophila Proteins Short linear motif Computer Simulation Immunophilins education Molecular Biology 030304 developmental biology 0303 health sciences education.field_of_study Microscopy Confocal biology Chemistry RNA Nuclear Proteins General Medicine liquid-liquid phase separation 021001 nanoscience & nanotechnology biology.organism_classification Intrinsically Disordered Proteins FKBP Drosophila melanogaster Microscopy Fluorescence Ribosome Subunits Spectrophotometry COS Cells Biophysics LLPS 0210 nano-technology Ribosomes Binding domain Protein Binding |
| Popis: | The FKBP39 from Drosophila melanogaster is a multifunctional regulatory immunophilin. It contains two globular domains linked by a highly charged disordered region. The N-terminal domain shows homology to the nucleoplasmin core domain, and the C-terminal domain is characteristic for the family of the FKBP immunophilin ligand binding domain. The specific partially disordered structure of the protein inspired us to investigate whether FKBP39 can drive spontaneous liquid-liquid phase separation (LLPS). Preliminary analyses using CatGranule and Pi-Pi contact predictors suggested a propensity for LLPS. Microscopy observations revealed that FKBP39 can self-concentrate to form liquid condensates. We also found that FKBP39 can lead to LLPS in the presence of RNA and peptides containing Arg-rich linear motifs derived from selected nuclear and nucleolar proteins. These heterotypic interactions have a stronger propensity for driving LLPS when compared to the interactions mediated by self-associating FKBP39 molecules. To investigate whether FKBP39 can drive LLPS in the cellular environment, we analysed it in fusion with YFP in COS-7 cells. The specific distribution and diffusion kinetics of FKBP39 examined by FRAP experiments provided evidence that immunophilin is an important driver of phase separation. The ability of FKBP39 to go into heterotypic interaction may be fundamental for ribosome subunits assembly. |
| Databáze: | OpenAIRE |
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