Linking amyloid protein aggregation and yeast survival.
Protein aggregation and amyloid formation lie behind an increasing number of human diseases. Here we describe the application of an "aggregation reporter", in which the test protein is fused to dihydrofolate reductase, as a general method to assess the intracellular solubility of amyloid proteins in eukaryotic background. Because the aggregation state of the target protein is linked directly to yeast cells survival in the presence of methotrexate, protein solubility can be monitored in vivo without the requirement of a functional assay for the protein of interest. In addition, the approach allows the in vivo visualization of the cellular location and aggregated state of the target protein. To demonstrate the applicability of the assay in the screening of genes or compounds that modulate amyloid protein aggregation in living cells, we have used as models the Alzheimer's amyloid β peptide, polyglutamine expansions of huntingtin, α-synuclein and non-aggregating variants thereof. Moreover, the anti-aggregational properties of small molecules and the effects of the yeast protein quality control machinery have also been evaluated using this method.
Departament de Bioquímica i Biologia Molecular, Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, E-08193 Bellaterra, Spain. salvador.ventura@uab.es
Montse
M
Morell
Natalia S
NS
de Groot
Josep
J
Vendrell
Francesc X
FX
Avilés
Salvador
S
Ventura
D016229:Amyloid beta-Peptides / Q000235:genetics* / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D050296:Microbial Viability; D018832:Molecular Chaperones / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000378:metabolism / Q000494:pharmacology; D012995:Solubility; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000378:metabolism
D016229:Amyloid beta-Peptides; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D054852:Small Molecule Libraries; D049452:Green Fluorescent Proteins; D013762:Tetrahydrofolate Dehydrogenase