AUTHOR=Danilov Lavrentii G. , Matveenko Andrew G. , Ryzhkova Varvara E. , Belousov Mikhail V. , Poleshchuk Olga I. , Likholetova Daria V. , Sokolov Petr A. , Kasyanenko Nina A. , Kajava Andrey V. , Zhouravleva Galina A. , Bondarev Stanislav A. TITLE=Design of a New [PSI+]-No-More Mutation in SUP35 With Strong Inhibitory Effect on the [PSI+] Prion Propagation JOURNAL=Frontiers in Molecular Neuroscience VOLUME=12 YEAR=2019 URL=https://www.frontiersin.org/journals/molecular-neuroscience/articles/10.3389/fnmol.2019.00274 DOI=10.3389/fnmol.2019.00274 ISSN=1662-5099 ABSTRACT=

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 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.