Yeasts as a Tool for Heterologous Gene Expression

Mokdad-Gargouri, Raja; Abdelmoula-Soussi, Salma; Hadiji-Abbes, Nadia; Yacoubi, Inès; Borchani-Chabchoub, Istabrak; Gargouri, Ali

doi:10.1007/978-1-61779-433-9_18

Cited by 24 publications

(19 citation statements)

References 23 publications

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“…After Bax, human p53 was the second heterologous proapoptotic gene that, although not present in the yeast genome, found to induce apoptosis‐like cell death when expressed in S. cerevisiae , thereby reinforcing the hypothesis that the apoptotic processes are conserved throughout the evolution from yeast to mammalian cells (Ligr et al ., ; Hadj Amor et al ., ; Greenwood & Ludovico, ). p53 induces yeast cell death with characteristic markers of apoptosis, such as DNA strand cleavage, the externalization of phosphatidylserines, and accumulation of ROS (Hadj Amor et al ., ; Mokdad‐Gargouri et al ., ). Very recently, it was reported that cell death induced by p53 expression in yeast is rather Yca1p independent and mainly mediated by Nuc1p (Palermo et al ., ), suggesting the importance of mitochondria in the p53‐induced cell death in yeast.…”

Section: P53mentioning

confidence: 97%

The expanding role of yeast in cancer research and diagnosis: insights into the function of the oncosuppressors p53 and BRCA1/2

et al. 2013

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When the glucose supply is high, despite the presence of oxygen, Saccharomyces cerevisiae uses fermentation as its main metabolic pathway and switches to oxidative metabolism only when this carbon source is limited. There are similarities between glucose-induced repression of oxidative metabolism of yeast and metabolic reprogramming of tumor cells. The glucose-induced repression of oxidative metabolism is regulated by oncogene homologues in yeast, such as RAS and Sch9p, the yeast homologue of Akt. Yeast also undergoes an apoptosis-like programmed cell death process sharing several features with mammalian apoptosis, including oxidative stress and a major role played by mitochondria. Evasion of apoptosis and sustained proliferative signaling are hallmarks of cancer. This, together with the possibility of heterologous expression of human genes in yeast, has allowed new insights to be obtained into the function of mammalian oncogenes/oncosuppressors. Here, we elaborate on the similarities between tumor and yeast cells underpinning the use of this model organism in cancer research. We also review the achievements obtained through heterologous expression in yeast of p53, BRCA1, and BRCA2, which are among the best-known cancer-susceptibility genes, with the aim of understanding their role in tumorigenesis. Yeast-cell-based functional assays for cancer genetic testing will also be dealt with.

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Section: P53mentioning

confidence: 97%

The expanding role of yeast in cancer research and diagnosis: insights into the function of the oncosuppressors p53 and BRCA1/2

et al. 2013

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“…Chemical inducers, such as IPTG, methanol or galactose, have been profusely employed for inducible gene regulation despite their obvious limitations including potential interference with metabolic processes, difficult removal from the culture media once added, and insufficient temporal and spatial/dose resolution (1). In addition, the cost of chemical inducers can restrict their uses in some industrial applications, whereas temperature induction or constitutive gene expression are chosen based on cost efficiency regardless of their far-from-optimal characteristics (2). Light constitutes a promissory alternative for the control of gene expression, considering its low cost, reduced toxic effects, adjustable levels, and high temporal and spatial resolution (3).…”

Section: Introductionmentioning

confidence: 99%

FUN-LOV: Fungal LOV domains for optogenetic control of gene expression and flocculation in yeast

Salinas

Rojas

Delgado

et al. 2018

Preprint

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Optogenetic switches permit accurate control of gene expression upon light stimulation. These synthetic switches have become a powerful tool for gene regulation, allowing modulation of customized phenotypes, overcoming the obstacles of chemical inducers and replacing their use by an inexpensive resource: light. In this work, we implemented FUN-LOV; an optogenetic switch based on the photon-regulated interaction of WC-1 and VVD, two LOV (Light Oxygen Voltage) blue-light photoreceptors from the fungus Neurospora crassa. When tested in yeast, FUN-LOV yields light-controlled gene expression with exquisite temporal resolution, and a broad dynamic range of over 1300-fold, as measured by a luciferase reporter. We also tested the FUN-LOV switch for heterologous protein expression in Saccharomyces cerevisiae, where Western blot analysis confirmed strong induction upon light stimulation, surpassing by 2.5 times the levels achieved with a classic GAL4/galactose chemical inducible system. Additionally, we utilized FUN-LOV to control the ability of yeast cells to flocculate. Light-controlled expression of the flocculin encoding gene FLO1, by the FUN-LOV switch, yielded Flocculation in Light (FIL), whereas the light-controlled expression of the co-repressor TUP1 provided Flocculation in Darkness (FID). Overall, the results revealed the potential of the FUN-LOV optogenetic switch to control two biotechnologically relevant phenotypes such as heterologous protein expression and flocculation, paving the road for the engineering of new yeast strains for industrial applications. Importantly, FUN-LOV’ s ability to accurately manipulate gene expression, with a high-temporal dynamic range, can be exploited in the analysis of diverse biological processes in various organisms.ImportanceOptogenetic switches are molecular devices which allow the control of different cellular processes by light, such as gene expression, providing a versatile alternative to chemical inducers. Herein, we report a novel optogenetic switch (FUN-LOV) based on the LOV-domain interaction of two blue-light photoreceptors (WC-1 and VVD) from the fungus N. crassa. In yeast cells, FUN-LOV allowed tight regulation of gene expression, with low background in darkness and a highly dynamic and potent control by light. We used FUN-LOV to optogenetically manipulate, in yeast, two biotechnologically relevant phenotypes: heterologous protein expression and flocculation, resulting in strains with potential industrial applications. Importantly, FUN-LOV can be implemented in diverse biological platforms to orthogonally control a multitude of cellular processes.

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“…S. cerevisiae SIC, generated using the pSynInsCPOT plasmid for increased insulin production when compared with the conventional strain, is a representative example [26,27]. Beyond S. cerevisiae, P. pastoris has been used to produce the human p53 tumor suppressor protein [28], and Schizosaccharomyces pombe has been widely used to produce hepatitis vaccines [24].…”

Section: Wine Beer and Breadmentioning

confidence: 99%

Anti-Contamination Strategies for Yeast Fermentations

Seo

Park

Jung³

et al. 2020

Microorganisms

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Yeasts are very useful microorganisms that are used in many industrial fermentation processes such as food and alcohol production. Microbial contamination of such processes is inevitable, since most of the fermentation substrates are not sterile. Contamination can cause a reduction of the final product concentration and render industrial yeast strains unable to be reused. Alternative approaches to controlling contamination, including the use of antibiotics, have been developed and proposed as solutions. However, more efficient and industry-friendly approaches are needed for use in industrial applications. This review covers: (i) general information about industrial uses of yeast fermentation, (ii) microbial contamination and its effects on yeast fermentation, and (iii) currently used and suggested approaches/strategies for controlling microbial contamination at the industrial and/or laboratory scale.

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Yeasts as a Tool for Heterologous Gene Expression

Cited by 24 publications

References 23 publications

The expanding role of yeast in cancer research and diagnosis: insights into the function of the oncosuppressors p53 and BRCA1/2

The expanding role of yeast in cancer research and diagnosis: insights into the function of the oncosuppressors p53 and BRCA1/2

FUN-LOV: Fungal LOV domains for optogenetic control of gene expression and flocculation in yeast

Anti-Contamination Strategies for Yeast Fermentations

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