Identification of PrP sequences essential for the interaction between the PrP polymers and Aβ peptide in a yeast-based assay

Rubel, Aleksandr A.; Ryzhova, Tatyana A.; Antonets, Kirill S.; Chernoff, Yury O.; Galkin, A. P.

doi:10.4161/pri.26867

Cited by 23 publications

(24 citation statements)

References 41 publications

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“…Therefore, we have also used the PrP90-231-GFP and Aβ1-42-GFP fusion proteins that are both produced at high levels in yeast cells (Fig S1A). The PrP-and Aβ-based constructs produced amyloid-like detergentresistant aggregates, in yeast, as shown previously [31] and confirmed by us (Fig S1B) using the semidenaturing agarose gel electrophoresis, SDD-AGE (see [32] (Fig 1A, B). This was in contrast to the yeast prionogenic QN-rich protein Lsb2 (fused to GFP), that promoted [PSI + ] formation in the presence of excess Sup35N (Fig 1A, B), as shown previously [23,33,34].…”

supporting

confidence: 87%

“…Then, respective chimeric genes were cut from plasmids pcDNA3.1(Zeo)-Sup35NM-NAC and pcDNA3.1(Zeo)-Sup35NM-IAPP with BamHI and XbaI, and inserted into the pmCUP1 vector at the position following the P CUP1 promoter. The plasmid coding for the C-terminal fusion of Lsb2 with GFP, expressed under under the P CUP1 promoter in the pRS316 backbone was constructed earlier [31]. The chimeric gene coding for Sup35N-LacZ was constructed by inserting the PCR amplified EcoRI-XbaI fragment that contains the lacZ coding sequence from the plasmid pSVA1 (kindly provided by Dr. M.D.…”

Section: Methodsmentioning

confidence: 99%

“…While several yeast assays for Aβ were proposed previously (e. g., [31,77,78,79]), none of them specifically address the initial nucleation of Aβ polymerization. Our approach could be used to map regions and identify amino acid residues specifically important for polymer nucleation, a crucial step triggering the subsequent amyloid formation and pathogenicity of Aβ, PrP, and other disease-related amyloidogenic proteins.…”

Section: Potential Applications Of the Yeast Prion Nucleation Assaymentioning

confidence: 99%

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Mammalian amyloidogenic proteins promote prion nucleation in yeast

Chandramowlishwaran

Sun

Casey

et al. 2018

Journal of Biological Chemistry

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Fibrous cross-β aggregates (amyloids) and their transmissible forms (prions) cause diseases in mammals (including humans) and control heritable traits in yeast. Initial nucleation of a yeast prion by transiently overproduced prion-forming protein or its (typically, QN-rich) prion domain is efficient only in the presence of another aggregated (in most cases, QN-rich) protein. Here, we demonstrate that a fusion of the prion domain of yeast protein Sup35 to some non-QN-rich mammalian proteins, associated with amyloid diseases, promotes nucleation of Sup35 prions in the absence of preexisting aggregates. In contrast, both a fusion of the Sup35 prion domain to a multimeric nonamyloidogenic protein, and an expression of a mammalian amyloidogenic protein that is not fused to the Sup35 prion domain failed to promote prion nucleation, further indicating that physical linkage of a mammalian amyloidogenic protein to the prion domain of a yeast protein is required for the nucleation of a yeast prion. Biochemical and cytological approaches confirmed the nucleation of protein aggregates in the yeast cell. Sequence alterations antagonizing or enhancing amyloidogenicity of human β amyloid (Aβ, associated with Alzheimer disease) and mouse PrP (associated with prion diseases) respectively antagonized or enhanced nucleation of a yeast prion by these proteins. The yeast-based prion nucleation assay, developed in our work, can be employed for mutational dissection of amyloidogenic proteins. We anticipate that it will aid in the identification of chemicals that influence initial amyloid nucleation and in searching for new amyloidogenic proteins in a variety of proteomes.

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supporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

Section: Potential Applications Of the Yeast Prion Nucleation Assaymentioning

confidence: 99%

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Mammalian amyloidogenic proteins promote prion nucleation in yeast

Chandramowlishwaran

Sun

Casey

et al. 2018

Journal of Biological Chemistry

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“…Notably, mammalian amyloidogenic proteins PrP (associated with transmissible spongiform encephalopathies) and Ab (associated with Alzheimer's disease) colocalize and even physically interact to each other (according to F€ orster resonance energy transfer, or FRET analysis) when coexpressed in yeast. 64 These proteins are also shown to interact to each other in mammalian and human brains [65][66][67] or in vitro, 68 confirming that coaggregation in yeast likely reflects actual physiological interactions.…”

Section: At Whichmentioning

confidence: 83%

Strain conformation controls the specificity of cross-species prion transmission in the yeast model

Grizel

Rubel

Chernoff

2016

Prion

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ABSTRACT. Transmissible self-assembled fibrous cross-b polymer infectious proteins (prions) cause neurodegenerative diseases in mammals and control non-Mendelian heritable traits in yeast. Cross-species prion transmission is frequently impaired, due to sequence differences in prionforming proteins. Recent studies of prion species barrier on the model of closely related yeast species show that colocalization of divergent proteins is not sufficient for the cross-species prion transmission, and that an identity of specific amino acid sequences and a type of prion conformational variant (strain) play a major role in the control of transmission specificity. In contrast, chemical compounds primarily influence transmission specificity via favoring certain strain conformations, while the species origin of the host cell has only a relatively minor input. Strain alterations may occur during cross-species prion conversion in some combinations. The model is discussed which suggests that different recipient proteins can acquire different spectra of prion strain conformations, which could be either compatible or incompatible with a particular donor strain.Correspondence to:

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“…Yeasts have also been used as a model system to unravel the molecular basis for several aging-related diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and cancer (Outeiro and Giorgini 2006 ;Rubel et al 2013 ;Willingham et al 2003 ). Moreover, with the large amount of data collected in yeast, systems biology approaches are easily applied to this organism for quantitative description of complex phenotypes such as aging (Lorenz et al 2009 ;Matecic et al 2010 ;Yizhak et al 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Aging in the Single-Celled Eukaryote, S. cerevisiae

Kennedy

2015

Stem Cell Aging: Mechanisms, Consequences, Rejuvenation

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In the last few decades, the budding yeast Saccharomyces cerevisiae has emerged as a simple and powerful model organism to study aging. Replicative aging and chronological aging are the two major models that have been established in yeast. In this chapter, we review the two aging model systems, focusing on genes and pathways that modulate replicative and chronological aging. The purpose of this chapter is to provide an overall understanding of the aging process in the single-celled yeast and a basis by which to generate models of molecular mechanisms that may affect aging stem cell populations in adult tissues, as well as the multicellular eukaryotes they inhabit.

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Identification of PrP sequences essential for the interaction between the PrP polymers and Aβ peptide in a yeast-based assay

Cited by 23 publications

References 41 publications

Mammalian amyloidogenic proteins promote prion nucleation in yeast

Mammalian amyloidogenic proteins promote prion nucleation in yeast

Strain conformation controls the specificity of cross-species prion transmission in the yeast model

Aging in the Single-Celled Eukaryote, S. cerevisiae

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