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Researchers inserted over 13,000 human genes into baker's yeast cells to study how proteins are packaged into extracellular vesicles (EVs), tiny membrane-bound particles that cells use to communicate. They successfully expressed 3,288 human proteins in yeast and identified 292 of these proteins in the EVs released by the yeast, with over 70% having yeast equivalents also found in natural yeast EVs. The findings suggest that the mechanisms controlling which proteins get sorted into EVs are evolutionarily conserved between yeast and humans.
Why it matters
This establishes yeast as a rapid, cost-effective model system for studying human cellular communication and EV biology. The approach could enable engineering of custom-designed EVs for drug delivery or therapeutic applications by exploiting conserved sorting mechanisms across species.
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⚠️ Preprint – Noch nicht peer-reviewed
Dieser Artikel wurde noch nicht von unabhängigen Experten begutachtet. Die Ergebnisse sind vorläufig und sollten mit Vorsicht interpretiert werden.
Extracellular vesicles (EVs) mediate intercellular communication by all organisms studied, from bacteria to yeast to man. Yet evolutionarily conserved mechanisms governing aspects of fundamental EV biology remain enigmatic. To address this, we sought to establish Saccharomyces cerevisiae (bakers yeast) as a model by identifying ectopically expressed human proteins sorted into yeast EVs. Using an optimized pooled cloning method, we inserted >13,000 human open reading frames (ORFs) upstream of an GFP tag within yeast expression plasmids. After transformation into S. cerevisiae, we confirmed expression of 3,288 GFP-tagged human proteins with diverse cellular expression levels and subcellular localizations. Heat stress triggered release of intact, lipid-bound, EGFP-positive small EVs from all transformant pools. Proteomic analysis identified 292 human proteins within EV samples, including canonical human EV biomarkers. Over 70% had yeast orthologs also found in yeast EVs suggesting conserved sorting mechanisms. Protein-protein interaction network analysis linked these EV cargoes to ESCRT-associated pathways. Finally, validation of seven candidates showed that DEF3A, ANXA2 and CLIC1 were enriched in yeast EVs. This study establishes an omics-compatible synthetic biology framework to humanize yeast EVs, begins to uncover conserved cargo sorting mechanisms, and supports future engineering of designer EVs.
Source: Human ORFeome expression in S. cerevisiae to better understand extracellular vesicle biology