Original Research ARTICLE
Design of a new [PSI+]-no-more mutation in SUP35 with strong inhibitory effect on the [PSI+] prion propagation
- 1Saint Petersburg State University, Russia
- 2All-Russian Research Institute of Agricultural Microbiology of the Russian Academy of Agricultural Sciences, Russia
- 3UMR5237 Centre de Recherche en Biologie cellulaire de Montpellier (CRBM), France
- 4Institut de Biologie Computationnelle, Université de Montpellier, France
A number of [PSI + ]-no-more (PNM) mutations, eliminating [PSI + ] prion, were previously described in SUP35. In this study, we designed and analyzed a new PNM mutation based on the parallel in-register β-structure of Sup35 prion fibrils suggested by the known experimental data. In such an arrangement, substitution of non-charged residues by charged ones may destabilize the fibril structure. We introduced Q33K/A34K amino acid substitutions into the Sup35 protein, corresponding allele was called sup35-M0. The mutagenized residues were chosen based on ArchCandy in silico prediction of high inhibitory effect on the amyloidogenic potential of Sup35. The experiments confirmed that Sup35-M0 leads to the elimination of [PSI + ] with high efficiency. Our data suggested that the elimination of the [PSI + ] prion is associated with the decreased aggregation properties of the protein. The new mutation can induce the prion with very low efficiency and is able to propagate only weak [PSI + ] prion variants. We also showed that Sup35-M0 protein co-aggregates with the wild-type Sup35 in vitro and in vivo. Moreover, our data confirmed the utility of the strategy of substitution of non-charged residues by charged ones to design new mutations to inhibit a prion formation.
Keywords: [PSI + ], Amyloid, ArchCandy, prion, Saccharomyces cerevisiae, SUP35 mutation, superpleated-β-structure
Received: 30 Aug 2019;
Accepted: 28 Oct 2019.
Copyright: © 2019 Danilov, Matveenko, Ryzhkova, Belousov, Poleshuk, Likholetova, Sokolov, Kasyanenko, Kajava, Zhouravleva and Bondarev. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Mx. Galina A. Zhouravleva, Saint Petersburg State University, Saint Petersburg, 199034, Saint Petersburg, Russia, firstname.lastname@example.org
Mx. Stanislav A. Bondarev, Saint Petersburg State University, Saint Petersburg, 199034, Saint Petersburg, Russia, email@example.com