Dynamic and Reversible Aggregation of the Human CAP Superfamily Member GAPR-1 in Protein Inclusions in Saccharomyces cerevisiae

JOURNAL OF MOLECULAR BIOLOGY

卷:433

Article

Sirati, Nafiseh¹  Popova, Blagovesta^2,3  Molenaar, Martijn R.^1,4  Verhoek, Iris C.¹  Braus, Gerhard H.^2,3  Kaloyanova, Dora, V¹  Helms, J. Bernd¹ 

[1] Univ Utrecht, Fac Vet Med, Div Cell Biol Metab & Canc, Utrecht, Netherlands

[2] Univ Gottingen, Dept Mol Microbiol & Genet, Inst Microbiol & Genet, Gottingen, Germany

[3] Univ Gottingen, Gottingen Ctr Mol Biosci GZMB, Inst Microbiol & Genet, Gottingen, Germany

[4] European Mol Biol Lab, Struct & Computat Biol Unit, Heidelberg, Germany

关键词: GLIPR-2;    condensates;    amyloids;    zinc;    myristoylation;   

DOI  :  10.1016/j.jmb.2021.167162

来源: Elsevier

【 摘 要 】

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 GolgiAssociated 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. (C) 2021 The Author(s). Published by Elsevier Ltd.

【 授权许可】

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