journal article
LitStream Collection
Li, Jiamei; Yang, Xuechen; Xu, Mingyue; Yin, Wenjing; Yang, Guoyu; Zheng, Yueting; Yang, Wei; Zhang, Wei
doi: 10.1002/yea.70005pmid: 41126396
The yeast Kluyveromyces marxianus (K. marxianus), characterized by its thermotolerance and rapid growth, is emerging as a promising new platform organism for the production of recombinant proteins. In this study, we constructed an expression vector designed for the efficient expression of exogenous proteins in K. marxianus. Initially, qPCR was employed to assess the expression efficiency of endogenous promoters within the yeast. The PDC1 promoter was selected, and its ability to drive the expression of EGFP was validated. The constructed vector exhibited high stability, maintaining approximately 5.2‐fold higher copy numbers than the K. marxianus genome after 72 hours of cultivation without hygromycin selection. Notably, the fluorescence signal intensity of K. marxianus harboring the vector was approximately 15.6‐fold higher than that of the wild‐type strain at 72 h. Subsequently, the cap gene of porcine circovirus type 3 (PCV3) was integrated into the vector, resulting in the production of soluble PCV3 cap protein. Electron microscopy analysis revealed that the PCV3 cap protein self‐assembled into virus‐like particles (VLPs). This study successfully established the expression vector and characterized its key elements in K. marxianus, which will facilitate further research on the expression of exogenous proteins in this yeast species. Moreover, the soluble expression of the PCV3 cap protein and its formation of VLPs provide a solid foundation for the future development of PCV3 vaccines.
Rocha, Miguel A.; Bhavani, Gowda; Fleischmann, Jacob
doi: 10.1002/yea.70007pmid: 41195863
Saccharomyces cerevisiae yeast cells have been shown to produce 18S and 25S ribosomal RNA molecules that are resistant to degradation by exonucleases, which require a 5′ monophosphate for activity. These resistant RNA species accumulate during the diauxic shift, a phase marked by reduced TOR signaling. To further investigate the link between TOR activity and the accumulation of resistant rRNA, we examined the effects of pharmacological TOR inhibition. Treatment with rapamycin, an active TOR suppressor, led to increased levels of resistant 18S and 25S RNA. Importantly, this accumulation was also observed in cells with constitutively active RNA polymerase I (CARA), indicating that the resistant RNA species arise independently of RNA Pol I transcriptional regulation. Similarly, a TOR1‐deleted mutant of Saccharomyces cerevisiae produces resistant 18S and 25S rRNA species in a sustained manner. Thiouracil labeling revealed that rRNA molecules generated during the logarithmic growth phase can be converted into the resistant form, suggesting a posttranscriptional modification process. Furthermore, thiouracil uptake assays demonstrated that overall rRNA synthesis decreases during the diauxic phase. Notably, decapping of the resistant rRNAs restored their sensitivity to exonucleases, indicating that the resistance is conferred by 5′ end modifications, likely involving the addition of one or more phosphate groups.
Ferreira‐Junior, Jose Ribamar; Lima Camandona, Vittoria; Barros, Mario H.
doi: 10.1002/yea.70008pmid: 41277874
Here, we review the use of Saccharomyces cerevisiae as a powerful model organism for studying cellular processes implicated in neurodegenerative disorders, including stress responses, proteostasis impairment, and vesicle trafficking defects. Over the last two decades, baker's yeast models have been developed for complex diseases such as Parkinson's, Alzheimer's, Huntington's, and Amyotrophic lateral sclerosis (ALS). Yeast cells expressing human proteins, such as amyloid‐β, α‐synuclein, huntingtin, and TDP‐43, have become crucial tools for high‐throughput drug screening aimed at counteracting disease progression. These yeast models have unveiled key components involved in the metabolism and toxicity of these proteins, enabling the identification of interacting partners and novel factors within each pathway. Importantly, these pathways were subsequently shown to be conserved in mammalian models. Furthermore, drug candidates identified using yeast models have provided significant leads for drug discovery, highlighting their potential for developing treatments for these neurodegenerative diseases.
doi: 10.1002/yea.70009pmid: 41445307
Yeasts have been intimately connected with human civilization for millennia, originally used for fermentation in food and beverage production. This article explores the multifaceted roles of yeasts—particularly Saccharomyces cerevisiae—as both a model organism and a cell factory. The historical journey of yeast research is chronicled from early fermentation practices to its central role in the molecular biology revolution. Notable discoveries using yeast have led to numerous Nobel Prizes, demonstrating its power in elucidating fundamental biological processes such as the eukaryal cell cycle, protein trafficking, transcription, and autophagy. The deep conservation of cellular pathways between yeast and humans, such as AMPK/Snf1 and TORC1/Tor1 signaling, further underscores yeast's value in biomedical research. Beyond its use in basic science, S. cerevisiae has become a preferred host for industrial biotechnology due to its genetic tractability, safety status, and ability to scale fermentation processes. Yeast has been engineered to produce a broad range of chemicals, fuels, and pharmaceuticals. Advanced tools in metabolic engineering—including genome‐scale metabolic models, multi‐omics analyses, and adaptive laboratory evolution—have driven remarkable improvements in yield, productivity, and strain robustness. These tools also offer insights into fundamental metabolic regulation and cellular adaptation. As the article discusses, yeast has not only illuminated the molecular workings of eukaryal life but also transformed industrial biotechnology. Its legacy and continued evolution affirm its indispensable role in science and technology.
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