scholarly journals Ultrastructural Analysis of Nanogold-labeled Endocytic Compartments of Yeast Saccharomyces cerevisiae Using a Cryosectioning Procedure

2009 ◽  
Vol 57 (8) ◽  
pp. 801-809 ◽  
Author(s):  
Janice Griffith ◽  
Fulvio Reggiori
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Model Organism ◽  
Specific Protein ◽  
Endosomal Trafficking ◽  
Protein Markers ◽  
Yeast Saccharomyces Cerevisiae ◽  
Novel Approach ◽  
Cargo Proteins ◽  
Endocytic Compartments ◽  
Endosomal System

Yeast Saccharomyces cerevisiae has been a valuable model organism for the study of the endosomal system of eukaryotic cells. Morphological analyses, however, have been limited because of the lack of specific protein markers and of procedures that lead to a satisfactory ultrastructural resolution. We have recently developed an immunoelectron microscopy (IEM) protocol adapted from the Tokuyasu method to prepare cryosections from mildly fixed yeast. This novel approach allows excellent cell preservation and a unique resolution of the yeast morphology. Here, we present a protocol that combines this procedure with the specific labeling of the various endosomal compartments with positively charged Nanogold. In particular, we show that this new protocol generates excellent results when applied for the examination of early and late endosomes, and of mutants with an endosomal trafficking defect. Importantly, this method is compatible with immunogold labeling of protein markers, and it is consequently appropriate for localization studies of both resident and cargo proteins. This new IEM protocol will be a valuable tool for the large community of scientists using yeast as a model system to investigate the membrane transport and the biogenesis of the endosomal system.

scholarly journals Maturation-driven transport and AP-1–dependent recycling of a secretory cargo in the Golgi

2019 ◽  
Vol 218 (5) ◽  
pp. 1582-1601 ◽  
Author(s):  
Jason C. Casler ◽  
Effrosyni Papanikou ◽  
Juan J. Barrero ◽  
Benjamin S. Glick
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fluorescence Imaging ◽  
Unexpected Finding ◽  
Maturation Process ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cargo Proteins

Golgi cisternal maturation has been visualized by fluorescence imaging of individual cisternae in the yeast Saccharomyces cerevisiae, but those experiments did not track passage of a secretory cargo. The expectation is that a secretory cargo will be continuously present within maturing cisternae as resident Golgi proteins arrive and depart. We tested this idea using a regulatable fluorescent secretory cargo that forms ER-localized aggregates, which dissociate into tetramers upon addition of a ligand. The solubilized tetramers rapidly exit the ER and then transit through early and late Golgi compartments before being secreted. Early Golgi cisternae form near the ER and become loaded with the secretory cargo. As predicted, cisternae contain the secretory cargo throughout the maturation process. An unexpected finding is that a burst of intra-Golgi recycling delivers additional secretory cargo molecules to cisternae during the early-to-late Golgi transition. This recycling requires the AP-1 adaptor, suggesting that AP-1 can recycle secretory cargo proteins as well as resident Golgi proteins.

scholarly journals Leveraging budding yeast Saccharomyces cerevisiae for discovering aging modulation substances for functional food

2019 ◽  
Vol 9 (5) ◽  
pp. 297
Author(s):  
Shaoyu Wang
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Bioactive Compounds ◽  
Functional Food ◽  
Budding Yeast ◽  
Model Organism ◽  
Cellular Systems ◽  
Cell Factory ◽  
Compression Of Morbidity ◽  
Bioactive Substances ◽  
Yeast Saccharomyces Cerevisiae

Background: Discovery of bioactive substances contained in functional food and the mechanism of their aging modulation are imperative steps in developing better, potent and safer functional food for promoting health and compression of morbidity in the aging population.  Budding yeast (Saccharomyces cerevisiae) is invaluable model organism for aging modulation and bioactive compounds discovery. In this paper we have conceptualised a framework for achieving such aim. This framework consists of four components: discovering targets for aging modulation, discovering and validating caloric restriction mimetics, acting as cellular systems for screening natural products or compounds for aging modulation and being a biological factory for producing bioactive compounds according to the roles the yeast systems play. It have been argued that the component of being a biological factory for producing bioactive compounds has much underexplored which also present an opportunity for new active substance discovery and validation for health promotion in functional food industry.Keywords: Aging modulation, budding yeast, functional food, bioactive substances, cell factory

scholarly journals The links between hypertrophy, reproductive potential and longevity in the Saccharomyces cerevisiae yeast.

2016 ◽  
Vol 63 (2) ◽  
Author(s):  
Mateusz Molon ◽  
Renata Zadrag-Tecza
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Volume ◽  
Genetic Background ◽  
Wild Type Strain ◽  
Reproductive Potential ◽  
Model Organism ◽  
Reproductive Capacity ◽  
Wild Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
Daughter Cells

The yeast Saccharomyces cerevisiae has long been used as a model organism for studying the basic mechanisms of aging. However, the main problem with the use of this unicellular fungus is the unit of "longevity". For all organisms, lifespan is expressed in units of time, while in the case of yeast it is defined by the number of daughter cells produced. Additionally, in yeast the phenotypic effects of mutations often show a clear dependence on the genetic background, suggesting the need for an analysis of strains representing different genetic backgrounds. Our results confirm the data presented in earlier papers that the reproductive potential is strongly associated with an increase in cell volume per generation. An excessive cell volume results in the loss of reproductive capacity. These data clearly support the hypertrophy hypothesis. The time of life of all analysed mutants, with the exception of sch9D, is the same as in the case of the wild-type strain. Interestingly, the 121% increase of the fob1D mutant's reproductive potential compared to the sfp1D mutant does not result in prolongation of the mutant's time of life (total lifespan).

scholarly journals Yeast as a Tool to Understand the Significance of Human Disease-Associated Gene Variants

Genes ◽  
2021 ◽  
Vol 12 (9) ◽  
pp. 1303
Author(s):  
Tiziana Cervelli ◽  
Alvaro Galli
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genetic Variability ◽  
Human Disease ◽  
Human Genetics ◽  
Model Organism ◽  
Model Organisms ◽  
Gene Variants ◽  
Yeast Saccharomyces Cerevisiae ◽  
Next Generation Dna Sequencing ◽  
Sequencing Technologies

At present, the great challenge in human genetics is to provide significance to the growing amount of human disease-associated gene variants identified by next generation DNA sequencing technologies. Increasing evidences suggest that model organisms are of pivotal importance to addressing this issue. Due to its genetic tractability, the yeast Saccharomyces cerevisiae represents a valuable model organism for understanding human genetic variability. In the present review, we show how S. cerevisiae has been used to study variants of genes involved in different diseases and in different pathways, highlighting the versatility of this model organism.

scholarly journals Whole-Genome Sequence and Variant Analysis of W303, a Widely-Used Strain of Saccharomyces cerevisiae

2017 ◽  
Vol 7 (7) ◽  
pp. 2219-2226 ◽  
Author(s):  
Kinnari Matheson ◽  
Lance Parsons ◽  
Alison Gammie
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genome Sequence ◽  
Model Organism ◽  
Laboratory Strain ◽  
Gene Families ◽  
Whole Genome Sequence ◽  
High Quality ◽  
Accurate Identification ◽  
Phenotypic Properties ◽  
Yeast Saccharomyces Cerevisiae

Abstract The yeast Saccharomyces cerevisiae has emerged as a superior model organism. Selection of distinct laboratory strains of S. cerevisiae with unique phenotypic properties, such as superior mating or sporulation efficiencies, has facilitated advancements in research. W303 is one such laboratory strain that is closely related to the first completely sequenced yeast strain, S288C. In this work, we provide a high-quality, annotated genome sequence for W303 for utilization in comparative analyses and genome-wide studies. Approximately 9500 variations exist between S288C and W303, affecting the protein sequences of ∼700 genes. A listing of the polymorphisms and divergent genes is provided for researchers interested in identifying the genetic basis for phenotypic differences between W303 and S288C. Several divergent functional gene families were identified, including flocculation and sporulation genes, likely representing selection for desirable laboratory phenotypes. Interestingly, remnants of ancestor wine strains were found on several chromosomes. Finally, as a test of the utility of the high-quality reference genome, variant mapping revealed more accurate identification of accumulated mutations in passaged mismatch repair-defective strains.

scholarly journals Effects of Non-Thermal Plasma on Yeast Saccharomyces cerevisiae

2021 ◽  
Vol 22 (5) ◽  
pp. 2247
Author(s):  
Peter Polčic ◽  
Zdenko Machala
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasma Cell ◽  
Cold Plasma ◽  
Cell Damage ◽  
Model Organism ◽  
Cell Interactions ◽  
Yeast Cells ◽  
Cell Physiology ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cold Plasmas

Cold plasmas generated by various electrical discharges can affect cell physiology or induce cell damage that may often result in the loss of viability. Many cold plasma-based technologies have emerged in recent years that are aimed at manipulating the cells within various environments or tissues. These include inactivation of microorganisms for the purpose of sterilization, food processing, induction of seeds germination, but also the treatment of cells in the therapy. Mechanisms that underlie the plasma-cell interactions are, however, still poorly understood. Dissection of cellular pathways or structures affected by plasma using simple eukaryotic models is therefore desirable. Yeast Saccharomyces cerevisiae is a traditional model organism with unprecedented impact on our knowledge of processes in eukaryotic cells. As such, it had been also employed in studies of plasma-cell interactions. This review focuses on the effects of cold plasma on yeast cells.

scholarly journals Signaling of Chloroquine-Induced Stress in the Yeast Saccharomyces cerevisiae Requires the Hog1 and Slt2 Mitogen-Activated Protein Kinase Pathways

2014 ◽  
Vol 58 (9) ◽  
pp. 5552-5566 ◽  
Author(s):  
Shivani Baranwal ◽  
Gajendra Kumar Azad ◽  
Vikash Singh ◽  
Raghuvir S. Tomar
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Histone Deacetylases ◽  
Model Organism ◽  
Biological Pathways ◽  
Human Cells ◽  
Mitogen Activated Protein Kinase ◽  
Yeast Saccharomyces Cerevisiae ◽  
Content Type ◽  
Mitogen Activated Protein ◽  
Glycerol 3 Phosphate Dehydrogenase

ABSTRACTChloroquine (CQ) has been under clinical use for several decades, and yet little is known about CQ sensing and signaling mechanisms or about their impact on various biological pathways. We employed the budding yeastSaccharomyces cerevisiaeas a model organism to study the pathways targeted by CQ. Our screening with yeast mutants revealed that it targets histone proteins and histone deacetylases (HDACs). Here, we also describe the novel role of mitogen-activated protein kinases Hog1 and Slt2, which aid in survival in the presence of CQ. Cells deficient in Hog1 or Slt2 are found to be CQ hypersensitive, and both proteins were phosphorylated in response to CQ exposure. CQ-activated Hog1p is translocated to the nucleus and facilitates the expression of GPD1 (glycerol-3-phosphate dehydrogenase), which is required for the synthesis of glycerol (one of the major osmolytes). Moreover, cells treated with CQ exhibited an increase in intracellular reactive oxygen species (ROS) levels and the effects were rescued by addition of reduced glutathione to the medium. The deletion of SOD1, the superoxide dismutase in yeast, resulted in hypersensitivity to CQ. We have also observed P38 as well as P42/44 phosphorylation in HEK293T human cells upon exposure to CQ, indicating that the kinds of responses generated in yeast and human cells are similar. In summary, our findings define the multiple biological pathways targeted by CQ that might be useful for understanding the toxicity modulated by this pharmacologically important molecule.

scholarly journals Studies on the Properties of the Sporulation Specific Protein Dit1 and Its Product Formyl Tyrosine

2020 ◽  
Vol 6 (2) ◽  
pp. 77
Author(s):  
Mostafa Basiony ◽  
Yan Yang ◽  
Guoyu Liu ◽  
Xiao-Dong Gao ◽  
Hideki Nakanishi
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Specific Protein ◽  
Spore Wall ◽  
Yeast Saccharomyces Cerevisiae ◽  
Crude Cell Lysate ◽  
Unknown Mechanism ◽  
Cell Lysate ◽  
Molecule Formation ◽  
Mild Hydrolysis ◽  
Mild Acid

The dityrosine layer is a unique structure present in the spore wall of the budding yeast Saccharomyces cerevisiae. The primary constituent of this layer is bisformyl dityrosine. A sporulation-specific protein, Dit1 is localized in the spore cytosol and produces a precursor of bisformyl dityrosine. Although Dit1 is similar to isocyanide synthases, the loss of Dit1 is not rescued by heterologous expression of the Pseudomonas aeruginosa isocyanide synthase, PvcA, indicating that Dit1 does not mediate isocyanidation. The product of Dit1 is most likely formyl tyrosine. Dit1 can produce its product when it is expressed in vegetative cells; however, formyl tyrosine was not detected in the crude cell lysate. We reasoned that formyl tyrosine is unstable and reacts with some molecule to form formyl tyrosine-containing molecules in the cell lysate. In support of this hypothesis, formyl tyrosine was detected when the lysate was hydrolyzed with a mild acid. The same property was also found for bisformyl dityrosine. Bisformyl dityrosine molecules assemble to form the dityrosine layer by an unknown mechanism. Given that bisformyl dityrosine can be released from the spore wall by mild hydrolysis, the process of formyl tyrosine-containing molecule formation may resemble the assembly of the dityrosine layer.

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