Dynamic and Reversible Aggregation of the Human CAP Superfamily Member GAPR-1 in Protein Inclusions in Saccharomyces cerevisiae
| Autor: | Sirati, Nafiseh, Popova, Blagovesta, Molenaar, Martijn R, Verhoek, Iris C, Braus, Gerhard H, Kaloyanova, Dora V, Helms, J Bernd, dB&C FR-RMSC RMSC, Veterinaire biochemie, dB&C FR-RMSC FR |
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| Přispěvatelé: | dB&C FR-RMSC RMSC, Veterinaire biochemie, dB&C FR-RMSC FR |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Cytoplasmic inclusion
Protein Conformation GLIPR-2 Saccharomyces cerevisiae Biophysics Protein degradation Crystallography X-Ray Protein Engineering 03 medical and health sciences Protein Aggregates Cytosol Protein Domains Structural Biology Organelle Humans amyloids Molecular Biology 030304 developmental biology Myristoylation Inclusion Bodies 0303 health sciences biology Chemistry myristoylation 030302 biochemistry & molecular biology zinc Membrane Proteins condensates Protein superfamily biology.organism_classification Cell biology Secretory protein Proteolysis |
| Zdroj: | Journal of Molecular Biology, 433(19), 1. Academic Press Inc. |
| ISSN: | 0022-2836 |
| Popis: | Many proteins that can assemble into higher order structures termed amyloids can also concentrate into cytoplasmic inclusions via liquid-liquid phase separation. Here, we study the assembly of human Golgi-Associated plant Pathogenesis Related protein 1 (GAPR-1), an amyloidogenic protein of the Cysteine-rich secretory proteins, Antigen 5, and Pathogenesis-related 1 proteins (CAP) protein superfamily, into cytosolic inclusions in Saccharomyces cerevisiae. Overexpression of GAPR-1-GFP results in the formation GAPR-1 oligomers and fluorescent inclusions in yeast cytosol. These cytosolic inclusions are dynamic and reversible organelles that gradually increase during time of overexpression and decrease after promoter shut-off. Inclusion formation is, however, a regulated process that is influenced by factors other than protein expression levels. We identified N-myristoylation of GAPR-1 as an important determinant at early stages of inclusion formation. In addition, mutations in the conserved metal-binding site (His54 and His103) enhanced inclusion formation, suggesting that these residues prevent uncontrolled protein sequestration. In agreement with this, we find that addition of Zn2+ metal ions enhances inclusion formation. Furthermore, Zn2+ reduces GAPR-1 protein degradation, which indicates stabilization of GAPR-1 in inclusions. We propose that the properties underlying both the amyloidogenic properties and the reversible sequestration of GAPR-1 into inclusions play a role in the biological function of GAPR-1 and other CAP family members. |
| Databáze: | OpenAIRE |
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