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Chandre Monerawela, Ursula Bond
Saccharomyces pastorianus is a recently evolved interspecies hybrid of Saccharomyces cerevisiae and Saccharomyces eubayanus used in the production of lager-type beers and has a long-standing history with the brewing industry. At least two distinct types of lager yeasts (Groups I and II) have been identified based on chromosome content and structure. One important feature of the genomes of lager yeasts is the presence of a set of hybrid chromosomes that emerged as a result of homeologous recombination events between the parental chromosomes...
August 8, 2017: Yeast
Albertin Warren, Chernova Maria, Durrens Pascal, Guichou Erwan, Sherman David, Masneuf-Pomarede Isabelle, Philippe Marullo
In the last two decades, the massive genome sequencing of strains belonging to the Saccharomyces genus revealed the complex reticulated evolution of this group. Among the various evolutionary mechanisms described, the introgression of large chromosomal regions resulting from interspecific hybridization has recently shed light on the Saccharomyces uvarum species. In this work we provide the de novo assembled genomes of four S. uvarum strains presenting more than 712 kb of introgressed loci inherited from both S...
August 5, 2017: Yeast
Gesa Zander, Wilfried Kramer, Anika Seel, Heike Krebber
Gle2/Rae1 is highly conserved from yeast to humans and described as an mRNA export factor. Additionally, it is implicated in the anaphase-promoting complex (APC)-mediated cell cycle regulation in higher eukaryotes. Here we identified an involvement for S. cerevisiae Gle2 in septin organization, which is crucial for cell cycle progression and cell division. Gle2 genetically and physically interacts with components of the septin-ring. Importantly, deletion of GLE2 leads to elongated buds, severe defects in septin-assembly and their cellular mislocalization...
August 4, 2017: Yeast
Jarkko Nikulin, Kristoffer Krogerus, Brian Gibson
The lager yeast hybrid (S. cerevisiae x S. eubayanus) possesses two key characteristics that are essential for lager brewing: efficient sugar utilization and cold tolerance. Here we explore the possibility that the lager yeast phenotype can be recreated by hybridizing S. cerevisiae ale yeast with a number of cold tolerant Saccharomyces species including S. arboricola, S. eubayanus, S. mikatae and S. uvarum. Interspecies hybrids performed better than parental strains in lager brewing conditions (12 °C and 12 °P wort), with the S...
July 28, 2017: Yeast
Lydia P Morris, Andrew B Conley, Natalya Degtyareva, I King Jordan, Paul W Doetsch
The DNA is cells is continuously exposed to reactive oxygen species resulting in toxic and mutagenic DNA damage. Although the repair of oxidative DNA damage occurs primarily through the base excision repair (BER) pathway, the nucleotide excision repair (NER) pathway processes some of the same lesions. In addition, damage tolerance mechanisms, such as recombination and translesion synthesis, enable cells to tolerate oxidative DNA damage, especially when BER and NER capacities are exceeded. Thus, disruption of BER alone or disruption of BER and NER in Saccharomyces cerevisiae leads to increased mutations as well as large-scale genomic rearrangements...
July 27, 2017: Yeast
Hua-Ping Pan, Ning Wang, Hiroyuki Tachikawa, Hideki Nakanishi, Xiao-Dong Gao
The yeast spore wall is an excellent model to study the assembly of an extracellular macromolecule structure. In the present study, mutants defective in ß-1,6-glucan synthesis, including kre1∆, kre6∆, kre9∆, and big1∆, were sporulated to analyze the effect of ß-1,6-glucan defects on the spore wall. Except for kre6∆, these mutant spores were sensitive to treatment with ether, suggesting that the mutations perturb the integrity of the spore wall. Morphologically, the mutant spores were indistinguishable from wild-type spores...
July 21, 2017: Yeast
Marianna Zambuto, Rossana Romaniello, Nicoletta Guaragnella, Patrizia Romano, Sonia Votta, Angela Capece
During active dry yeast (ADY) production process, cells are exposed to multiple stresses, such as thermal, oxidative and hyperosmotic shock. Previously, by analyzing cells in exponential growth phase, we selected an indigenous Saccharomyces cerevisiae wine strain, namely CD-6Sc, for its higher tolerance to desiccation and higher expression of specific desiccation stress-related genes in comparison to other yeast strains. In this study, we performed a desiccation treatment on stationary phase cells by comparing the efficacy of two different methods: a "laboratory dry test" in a small-scale (mild stress) and a treatment by spray-drying (severe stress), one of the most appropriate preservation method for yeasts and other micro-organisms...
July 21, 2017: Yeast
Dominika M Wloch-Salamon, Roberta M Fisher, Birgitte Regenberg
Division of labour between different specialized cell types is a central part of how we describe complexity in multicellular organisms. However, it is increasingly being recognised that division of labour also plays an important role in the lives of predominantly unicellular organisms. Saccharomyces cerevisiae displays several phenotypes that could be considered a division of labour, including quiescence, apoptosis and biofilm formation, but they have not been explicitly treated as such. We discuss each of these examples, using a definition of division of labour that involves phenotypic variation between cells within a population, cooperation between cells performing different tasks, and maximisation of the inclusive fitness of all cells involved...
July 5, 2017: Yeast
Verónica Mixão, Toni Gabaldón
Hybridization between different species can result in the emergence of new lineages and adaptive phenotypes. Occasionally, hybridization in fungal organisms can drive the appearance of opportunistic lifestyles or shifts to new hosts, resulting in the emergence of novel pathogens. In recent years, an increasing number of studies have documented the existence of hybrids in diverse yeast clades, including some comprising human pathogens. Comparative and population genomics studies performed on these clades are enabling us to understand what roles hybridization may play in the evolution and emergence of a virulence potential towards humans...
July 5, 2017: Yeast
Carolina Camelo, Filipe Vilas-Boas, Andreia Pereira Cepeda, Carla Real, Joana Barros-Martins, Francisco Pinto, Helena Soares, H Susana Marinho, Luisa Cyrne
During exposure of yeast cells to low levels of hydrogen peroxide (H2 O2 ), the expression of several genes is regulated for cells to adapt to the surrounding oxidative environment. Such adaptation involves modification of plasma membrane lipid composition, reorganization of ergosterol-rich microdomains and altered gene expression of proteins involved in lipid and vesicle traffic, to decrease permeability to exogenous H2 O2 . Opi1p is a transcriptional repressor that is inactive when present at the nuclear membrane/endoplasmic reticulum, but represseses transcription of inositol upstream activating sequence (UASINO )-containing genes, many of which are involved in the synthesis of phospholipids and fatty acids, when it is translocated to the nucleus...
June 5, 2017: Yeast
Su Jung Lee, Rashmi Ramesh, Valerie de Boor, Jan M Gebler, Richard C Silva, Evelyn Sattlegger
The common method for liberating proteins from Saccharomyces cerevisiae cells involves mechanical cell disruption using glass beads and buffer containing inhibitors (protease, phosphatase, and/or kinase inhibitors), followed by centrifugation to remove cell debris. This procedure requires the use of costly inhibitors, and is laborious, in particular when many samples need to be processed. Also, enzymatic reactions can still occur during harvesting and cell breakage. As a result low abundant and labile proteins may be degraded, and enzymes such as kinases and phosphatases may still modify proteins during and after cell lysis...
June 1, 2017: Yeast
Nurzhan Kuanyshev, Giusy M Adamo, Danilo Porro, Paola Branduardi
Zygosaccharomyces bailii is a non-Saccharomyces budding yeast known as one of the most aggressive food spoilage microorganism, often isolated as contaminant during wine fermentation, as well as from many acidic, high sugar and canned foods. The spoilage ability relies on the yeast's unique feature to tolerate the most common preservatives such as sulfite, dimethyl dicarbonate, acetic acid, and sorbic acid. Therefore, many studies focused on the description of this peculiar tolerance with the aim of developing preventative measures against Z...
May 27, 2017: Yeast
Xinping Lin, Ning Gao, Sasa Liu, Sufang Zhang, Shuang Song, Chaofan Ji, Xiuping Dong, Yichen Su, Zongbao Kent Zhao, Beiwei Zhu
The red yeast Rhodosporidium toruloides is a known lipid producer capable of accumulating large amounts of triacylglycerols and carotenoids. However, it remains challenging to study its carotenoid production profiles owing to limited biochemical information and inefficient genetic tools. Here we used an Agrobacterium tumefaciens-mediated transformation (ATMT) to change its carotenoid production and profiles. We constructed R. toruloides NP11 mutant libraries with ATMT, selected three mutants with different colours, characterized their carotenoid products by high-pressure liquid chromatography-mass spectrometry (HPLC-MS) analysis and assured differences among those strains in terms of carotenoid production and its composition profiles...
August 2017: Yeast
Céline Callens, Nelson C Coelho, Aaron W Miller, Maria Rosa Domingo Sananes, Maitreya J Dunham, Matthieu Denoual, Damien Coudreuse
Maintenance of long-term cultures of yeast cells is central to a broad range of investigations, from metabolic studies to laboratory evolution assays. However, repeated dilutions of batch cultures lead to variations in medium composition, with implications for cell physiology. In Saccharomyces cerevisiae, powerful miniaturized chemostat setups, or ministat arrays, have been shown to allow for constant dilution of multiple independent cultures. Here we set out to adapt these arrays for continuous culture of a morphologically and physiologically distinct yeast, the fission yeast Schizosaccharomyces pombe, with the goal of maintaining constant population density over time...
August 2017: Yeast
Makoto J Ohira, David G Hendrickson, R Scott McIsaac, Nicholas Rhind
The fission yeast Schizosaccharomyces pombe lacks a diverse toolkit of inducible promoters for experimental manipulation. Available inducible promoters suffer from slow induction kinetics, limited control of expression levels and/or a requirement for defined growth medium. In particular, no S. pombe inducible promoter systems exhibit a linear dose-response, which would allow expression to be tuned to specific levels. We have adapted a fast, orthogonal promoter system with a large dynamic range and a linear dose response, based on β-estradiol-regulated function of the human oestrogen receptor, for use in S...
August 2017: Yeast
Ruben Hohnholz, Kim Julia Pohlmann, Tilman Achstetter
Yeast episomal shuttle vectors (YEp type) are commonly used in fundamental research and biotechnology whenever elevated product levels are desired. Their instability, however, poses an impediment not only in industrial scale fermentation. In order to analyse instability which might be linked to plasmid structure, a series of YEp type plasmids that are identical in size has been assembled, differing only in the overall arrangement of the fragments used. The performance of the eight plasmid isoforms was studied with respect to mitotic stability...
June 2017: Yeast
Bethany Nancolas, Ian D Bull, Richard Stenner, Virginie Dufour, Paul Curnow
The alcohol-O-acyltransferases are bisubstrate enzymes that catalyse the transfer of acyl chains from an acyl-coenzyme A (CoA) donor to an acceptor alcohol. In the industrial yeast Saccharomyces cerevisiae this reaction produces acyl esters that are an important influence on the flavour of fermented beverages and foods. There is also a growing interest in using acyltransferases to produce bulk quantities of acyl esters in engineered microbial cell factories. However, the structure and function of the alcohol-O-acyltransferases remain only partly understood...
June 2017: Yeast
Robert Gnügge, Fabian Rudolf
Yeast shuttle vectors are indispensable tools in yeast research. They enable cloning of defined DNA sequences in Escherichia coli and their direct transfer into Saccharomyces cerevisiae cells. There are three types of commonly used yeast shuttle vectors: centromeric plasmids, episomal plasmids and integrating plasmids. In this review, we discuss the different plasmid systems and their characteristic features. We focus on their segregational stability and copy number and indicate how to modify these properties...
May 2017: Yeast
Yueping Zhang, Nina D Serratore, Scott D Briggs
PCR-mediated homologous recombination is a powerful approach to introduce epitope tags into the chromosomal loci at the N-terminus or the C-terminus of targeted genes. Although strategies of C-terminal epitope tagging of target genes at their loci are simple and widely used in yeast, C-terminal epitope tagging is not practical for all proteins. For example, a C-terminal tag may affect protein function or a protein may get cleaved or processed, resulting in the loss of the epitope tag. Therefore, N-terminal epitope tagging may be necessary to resolve these problems...
May 2017: Yeast
Alexander Lorenz
DNA helicases are ATP-driven motor proteins which translocate along DNA capable of dismantling DNA-DNA interactions and/or removing proteins bound to DNA. These biochemical capabilities make DNA helicases main regulators of crucial DNA metabolic processes, including DNA replication, DNA repair, and genetic recombination. This budding topic will focus on reviewing the function of DNA helicases important for homologous recombination during meiosis, and discuss recent advances in how these modulators of meiotic recombination are themselves regulated...
May 2017: Yeast
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