Prion Replication in the Mammalian Cytosol: Functional Regions within a Prion Domain Driving Induction, Propagation, and Inheritance

Duernberger, Yvonne; Liu, Shu; Riemschoss, Katrin; Paulsen, Lydia; Bester, Romina; Kuhn, Peer‐Hendrik; Schölling, Manuel; Lichtenthaler, Stefan F.; Vorberg, Ina

doi:10.1128/mcb.00111-18

Cited by 15 publications

(14 citation statements)

References 89 publications

(149 reference statements)

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“…Thus, independent of the fibrillization protocol, only p103-113 repeatedly induced NM-GFP agg in recipient cells, consistent with the in vitro results described above. This is in line with our recent finding demonstrating that amino acid residues 98-123 serve as a nucleation core in cytosolically expressed Sup35 NM that drives prion induction in mammalian N2a cells (Duernberger et al, 2018). Importantly, none of the fibrillized peptides used in this study show cytotoxic effects in mouse neuroblastoma cells ( Figure S2).…”

Section: Induction Of Cytosolic Sup35 Nm Aggregation By Synthetic P10supporting

confidence: 92%

“…These p103-113-induced aggregates indeed possess archetypal prion properties in mammalian cells, including inheritance of aggregates by progeny and cell-to-cell spreading propensity. Interestingly, deletion studies of Sup35 NM expressed in N2a cells previously demonstrated that a region comprising amino acid residues 98-123 is critically involved in prion induction and maintenance, arguing that this region represents the nucleation core of the NM prion state in a mammalian cell environment (Duernberger et al, 2018). The fact that other studies (Osherovich et al, 2004;Shkundina et al, 2006) showed that this region is dispensable for prion activity in yeast cells highlights that, depending on the host, different regions can nucleate and/or seed Sup35 NM prions.…”

Section: Discussionmentioning

confidence: 99%

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Entropic Bristles Tune the Seeding Efficiency of Prion-Nucleating Fragments

et al. 2020

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Section: Induction Of Cytosolic Sup35 Nm Aggregation By Synthetic P10supporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Entropic Bristles Tune the Seeding Efficiency of Prion-Nucleating Fragments

et al. 2020

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“…Interestingly, the prion-like behavior is not restricted to the original host. For example, cell-to-cell propagation of amyloid aggregates has been successfully reported in mammalian cells for the NM prion domain of the Saccharomyces cerevisiae translation termination factor Sup35 (24)(25)(26)(27)(28). Sup35-NM can also propagate in bacteria, provided that a second specific prion-inducing amyloid required for the prionization of Sup35 in S. cerevisiae is also expressed in the recipient cells (29).…”

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confidence: 99%

Intercellular Transmission of a Synthetic Bacterial Cytotoxic Prion-Like Protein in Mammalian Cells

Revilla-García

Fernández

Álamo

et al. 2020

mBio

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RepA is a bacterial protein that builds intracellular amyloid oligomers acting as inhibitory complexes of plasmid DNA replication. When carrying a mutation enhancing its amyloidogenesis (A31V), the N-terminal domain (WH1) generates cytosolic amyloid particles that are inheritable within a bacterial lineage. Such amyloids trigger in bacteria a lethal cascade reminiscent of mitochondrial impairment in human cells affected by neurodegeneration. To fulfill all the criteria to qualify as a prion-like protein, horizontal (intercellular) transmissibility remains to be demonstrated for RepA-WH1. Since this is experimentally intractable in bacteria, here we transiently expressed in a murine neuroblastoma cell line the soluble, barely cytotoxic RepA-WH1 wild type [RepA-WH1(WT)] and assayed its response to exposure to in vitro-assembled RepA-WH1(A31V) amyloid fibers. In parallel, murine cells releasing RepA-WH1(A31V) aggregates were cocultured with human neuroblastoma cells expressing RepA-WH1(WT). Both the assembled fibers and donor-derived RepA-WH1(A31V) aggregates induced, in the cytosol of recipient cells, the formation of cytotoxic amyloid particles. Mass spectrometry analyses of the proteomes of both types of injured cells pointed to alterations in mitochondria, protein quality triage, signaling, and intracellular traffic. Thus, a synthetic prion-like protein can be propagated to, and become cytotoxic to, cells of organisms placed at such distant branches of the tree of life as bacteria and mammalia, suggesting that mechanisms of protein aggregate spreading and toxicity follow default pathways. IMPORTANCE Proteotoxic amyloid seeds can be transmitted between mammalian cells, arguing that the intercellular exchange of prion-like protein aggregates can be a common phenomenon. RepA-WH1 is derived from a bacterial intracellular functional amyloid protein, engineered to become cytotoxic in Escherichia coli. Here, we have studied if such bacterial aggregates can also be transmitted to, and become cytotoxic to, mammalian cells. We demonstrate that RepA-WH1 is capable of entering naive cells, thereby inducing the cytotoxic aggregation of a soluble RepA-WH1 variant expressed in the cytosol, following the same trend that had been described in bacteria. These findings highlight the universality of one of the central principles underlying prion biology: No matter the biological origin of a given prion-like protein, it can be transmitted to a phylogenetically unrelated recipient cell, provided that the latter expresses a soluble protein onto which the incoming protein can readily template its amyloid conformation.

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“…Since there is obvious resemblance between its proposed mechanism of propagation and that of the mammalian prion disease, Sup35 is often used as a model protein to study the prion disease 19,20 . It has been shown that Sup35 may propagate as a prion in mammalian cells and that GPI anchoring facilitates aggregate propagation between N2a cells, resembling mammalian prion behavior [21][22][23][24] .…”

Section: Introductionmentioning

confidence: 99%

Membrane composition and lipid to protein ratio modulate amyloid kinetics of yeast prion protein

Bandyopadhyay

Sannigrahi

Chattopadhyay

2020

Preprint

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Understanding of prion aggregation in membrane environment may help to ameliorate neurodegenerative complications caused by the amyloid forms of prions. Here, we investigated the membrane binding induced aggregation of yeast prion protein Sup35. Using the combination of fluorescence correlation spectroscopy (FCS) at single molecule resolution and other biophysical studies, we establish that lipid composition and lipid/protein ratio are key modulators of the aggregation kinetics of Sup35. In the presence of zwitterionic membrane, Sup35 exhibited a novel biphasic aggregation kinetics at lipid/protein ratio ranging between 20:1 and 70:1 (termed here as the Optimum Lipid Concentration, OLC). In ratios below (Low Lipid Concentration, LLC) and above (ELC, Excess Lipid Concentration) that range, the aggregation was found to be monophasic. In contrast, in the presence of negatively charged membrane, we did not observe any bi-phasic aggregation kinetics in the entire range of protein to lipid ratios. The toxicity of the aggregates formed within OLC range was found to be greater. Our results provide a mechanistic description of the role that membrane-concentration/composition-modulated-aggregation may play in neurodegenerative diseases.

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Prion Replication in the Mammalian Cytosol: Functional Regions within a Prion Domain Driving Induction, Propagation, and Inheritance

Cited by 15 publications

References 89 publications

Entropic Bristles Tune the Seeding Efficiency of Prion-Nucleating Fragments

Entropic Bristles Tune the Seeding Efficiency of Prion-Nucleating Fragments

Intercellular Transmission of a Synthetic Bacterial Cytotoxic Prion-Like Protein in Mammalian Cells

Membrane composition and lipid to protein ratio modulate amyloid kinetics of yeast prion protein

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