English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/116705
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL
Exportar a otros formatos:
Title

Engineered bacterial hydrophobic oligopeptide repeats in a synthetic yeast prion, [REP-PSI+]

AuthorsGasset-Rosa, F. ; Giraldo, R.
KeywordsAmyloidcross-seeding
Prionvariants/strains
RepA-WH1prionoid
[REP-PSI+] prion
Saccharomyces cerevisiae
Syntheticbiology
Issue Date21-Apr-2015
PublisherFrontiers Media
CitationFront. Microbiol.6:311
AbstractThe yeast translation termination factor Sup35p, by aggregating as the [PSI (+)] prion, enables ribosomes to read-through stop codons, thus expanding the diversity of the Saccharomyces cerevisiae proteome. Yeast prions are functional amyloids that replicate by templating their conformation on native protein molecules, then assembling as large aggregates and fibers. Prions propagate epigenetically from mother to daughter cells by fragmentation of such assemblies. In the N-terminal prion-forming domain, Sup35p has glutamine/asparagine-rich oligopeptide repeats (OPRs), which enable propagation through chaperone-elicited shearing. We have engineered chimeras by replacing the polar OPRs in Sup35p by up to five repeats of a hydrophobic amyloidogenic sequence from the synthetic bacterial prionoid RepA-WH1. The resulting hybrid, [REP-PSI (+)], (i) was functional in a stop codon read-through assay in S. cerevisiae; (ii) generates weak phenotypic variants upon both its expression or transformation into [psi (-)] cells; (iii) these variants correlated with high molecular weight aggregates resistant to SDS during electrophoresis; and (iv) according to fluorescence microscopy, the fusion of the prion domains from the engineered chimeras to the reporter protein mCherry generated perivacuolar aggregate foci in yeast cells. All these are signatures of bona fide yeast prions. As assessed through biophysical approaches, the chimeras assembled as oligomers rather than as the fibers characteristic of [PSI (+)]. These results suggest that it is the balance between polar and hydrophobic residues in OPRs what determines prion conformational dynamics. In addition, our findings illustrate the feasibility of enabling new propagation traits in yeast prions by engineering OPRs with heterologous amyloidogenic sequence repeats.
Description12 p.-5 fig. [This Document is Protected by copyright and was first published by Frontiers. All rights reserved. it is reproduced with permission.]
Publisher version (URL)http://dx.doi.org/ 10.3389/fmicb.2015.00311
URIhttp://hdl.handle.net/10261/116705
DOI10.3389/fmicb.2015.00311
ISSN1664-302X
E-ISSN1664-302X
Appears in Collections:(CIB) Artículos
Files in This Item:
File Description SizeFormat 
Frontiers in Microbiology_R. Giraldo_2015,.pdf4,36 MBAdobe PDFThumbnail
View/Open
Show full item record
Review this work
 

Related articles:


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.