Assays to Monitor Autophagy in Saccharomyces cerevisiae

Torggler, Raffaela; Papinski, Daniel; Kraft, Claudine

doi:10.3390/cells6030023

Cited by 55 publications

(41 citation statements)

References 119 publications

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“…On the other hand, Bae and colleagues have declared TCTP a negative regulator of basal and rapamycin-induced non-selective autophagy [11]. Since TCTP is an evolutionarily highly conserved protein, we used a pioneer model organism for studying autophagy, budding yeast Saccharomyces cerevisiae [12], reviewed in [13][14][15] to test the effect of yeast TCTP on autophagy. The S. cerevisiae is usually batch cultured and its growth in the culture is highly affected by a carbon source.…”

Section: Introductionmentioning

confidence: 99%

Mmi1, the Yeast Ortholog of Mammalian Translationally Controlled Tumor Protein (TCTP), Negatively Affects Rapamycin-Induced Autophagy in Post-Diauxic Growth Phase

Vojtová¹,

Hašek²

2020

Cells

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Translationally controlled tumor protein (TCTP) is a multifunctional and highly conserved protein from yeast to humans. Recently, its role in non-selective autophagy has been reported with controversial results in mammalian and human cells. Herein we examine the effect of Mmi1, the yeast ortholog of TCTP, on non-selective autophagy in budding yeast Saccharomyces cerevisiae, a well-established model system to monitor autophagy. We induced autophagy by nitrogen starvation or rapamycin addition and measured autophagy by using the Pho8Δ60 and GFP-Atg8 processing assays in WT, mmi1Δ, and in autophagy-deficient strains atg8Δ or atg1Δ. Our results demonstrate that Mmi1 does not affect basal or nitrogen starvation-induced autophagy. However, an increased rapamycin-induced autophagy is detected in mmi1Δ strain when the cells enter the post-diauxic growth phase, and this phenotype can be rescued by inserted wild-type MMI1 gene. Further, the mmi1Δ cells exhibit significantly lower amounts of reactive oxygen species (ROS) in the post-diauxic growth phase compared to WT cells. In summary, our study suggests that Mmi1 negatively affects rapamycin-induced autophagy in the post-diauxic growth phase and supports the role of Mmi1/TCTP as a negative autophagy regulator in eukaryotic cells.

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Section: Introductionmentioning

confidence: 99%

Mmi1, the Yeast Ortholog of Mammalian Translationally Controlled Tumor Protein (TCTP), Negatively Affects Rapamycin-Induced Autophagy in Post-Diauxic Growth Phase

Vojtová¹,

Hašek²

2020

Cells

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“…The mechanisms of these initial stages have been intensively studied and described in numerous review papers, e.g., [30][31][32][33]. In yeast and plants, the autophagosome fuses with the vacuole creating an autophagic body that is quickly degraded by vacuolar lytic enzymes [34][35][36][37]. In animals, the autophagic body is not formed because autophagosome fuses with the lysosome, which delivers lytic enzymes enabling the degradation of the cargo inside the autolysosome (Figure 1) [29,38].…”

Section: Formation and Trafficking Of Autophagosomementioning

confidence: 99%

“…Proteins involved in the degradation of the autophagic body in yeast are proteinase A (Pep4) and proteinase B (Prb1) (Figure 2, Table 3). These proteins are involved in the activation of cascades of other vacuolar proteases and hydrolases, which are indicated as key factors involved in the degradation of the autophagic body in yeast [34,115,116]. However, the cascades involved in autophagic body degradation have not yet been identified in detail.…”

Section: Degradation Of the Autophagic Body And Metabolite Efflux Fromentioning

confidence: 99%

“…The best-known and described protein involved in the degradation of the autophagic body in yeast is Atg15, a putative lipase (Figure 2, Table 3). It has been proven that this protein plays a role in the degradation of the autophagic body, not only through the decomposition and recycling of components of the autophagic body membrane but also due to it being a key protein involved in the degradation of the cargo located inside the autophagic body [34,[117][118][119][120]. In addition to those described above, there are several other proteins that are thought to be involved in the degradation of the autophagic body in yeast, but their mechanism of action and functions are not yet fully understood.…”

Section: Degradation Of the Autophagic Body And Metabolite Efflux Fromentioning

confidence: 99%

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Completing Autophagy: Formation and Degradation of the Autophagic Body and Metabolite Salvage in Plants

Stefaniak

Wojtyla

Pietrowska‐Borek

et al. 2020

IJMS

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Autophagy is an evolutionarily conserved process that occurs in yeast, plants, and animals. Despite many years of research, some aspects of autophagy are still not fully explained. This mostly concerns the final stages of autophagy, which have not received as much interest from the scientific community as the initial stages of this process. The final stages of autophagy that we take into consideration in this review include the formation and degradation of the autophagic bodies as well as the efflux of metabolites from the vacuole to the cytoplasm. The autophagic bodies are formed through the fusion of an autophagosome and vacuole during macroautophagy and by vacuolar membrane invagination or protrusion during microautophagy. Then they are rapidly degraded by vacuolar lytic enzymes, and products of the degradation are reused. In this paper, we summarize the available information on the trafficking of the autophagosome towards the vacuole, the fusion of the autophagosome with the vacuole, the formation and decomposition of autophagic bodies inside the vacuole, and the efflux of metabolites to the cytoplasm. Special attention is given to the formation and degradation of autophagic bodies and metabolite salvage in plant cells.

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“…These authors reported that autophagy takes place during secondary fermentation of sparkling wines and proposed the use of yeast strains with deregulated autophagy in order to accelerate the autolysis process. Autophagy is induced mainly under starvation conditions and involves the transport and degradation of cytoplasmic compounds in the vacuole [7,8]. It can be classified into two main types: macroautophagy and microautophagy, which are both selective and non-selective processes.…”

Section: Introductionmentioning

confidence: 99%

Autophagic Proteome in Two Saccharomyces cerevisiae Strains during Second Fermentation for Sparkling Wine Elaboration

et al. 2020

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A correlation between autophagy and autolysis has been proposed in order to accelerate the acquisition of wine organoleptic properties during sparkling wine elaboration. In this context, a proteomic analysis was carried out in two industrial Saccharomyces cerevisiae strains (P29, conventional sparkling wine strain and G1, implicated in sherry wine elaboration) with the aim of studying the autophagy-related proteome and comparing the effect of CO2 overpressure during sparkling wine elaboration. In general, a detrimental effect of pressure and second fermentation development on autophagy-related proteome was observed in both strains, although it was more pronounced in flor yeast strain G1. Proteins mainly involved in autophagy regulation and autophagosome formation in flor yeast G1, and those required for vesicle nucleation and expansion in P29 strain, highlighted in sealed bottle. Proteins Sec2 and Sec18 were detected 3-fold under pressure conditions in P29 and G1 strains, respectively. Moreover, ‘fingerprinting’ obtained from multivariate data analysis established differences in autophagy-related proteome between strains and conditions. Further research is needed to achieve more solid conclusions and design strategies to promote autophagy for an accelerated autolysis, thus reducing cost and time production, as well as acquisition of good organoleptic properties.

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Assays to Monitor Autophagy in Saccharomyces cerevisiae

Cited by 55 publications

References 119 publications

Mmi1, the Yeast Ortholog of Mammalian Translationally Controlled Tumor Protein (TCTP), Negatively Affects Rapamycin-Induced Autophagy in Post-Diauxic Growth Phase

Mmi1, the Yeast Ortholog of Mammalian Translationally Controlled Tumor Protein (TCTP), Negatively Affects Rapamycin-Induced Autophagy in Post-Diauxic Growth Phase

Completing Autophagy: Formation and Degradation of the Autophagic Body and Metabolite Salvage in Plants

Autophagic Proteome in Two Saccharomyces cerevisiae Strains during Second Fermentation for Sparkling Wine Elaboration

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