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Ruckenstuhl C, Büttner S, Carmona-Gutierrez D, Eisenberg T, Kroemer G, Sigrist SJ, Fröhlich KU, Madeo F PLoS ONE. 2009;4(2):e4592 BACKGROUND: Otto Warburg observed that cancer cells are often characterized by intense glycolysis in the presence of oxygen and a concomitant decrease in mitochondrial respiration. Research has mainly focused on a possible connection between increased glycolysis and tumor development whereas decreased respiration has largely been left unattended. Therefore, a causal relation between decreased respiration and tumorigenesis has not been demonstrated. METHODOLOGY/PRINCIPAL FINDINGS: For this purpose, colonies of Saccharomyces cerevisiae, which is suitable for manipulation of mitochondrial respiration and shows mitochondria-mediated cell death, were used as a model. Repression of respiration as well as ROS-scavenging via glutathione inhibited apoptosis and conferred a survival advantage during seeding and early development of this fast proliferating solid cell population. In contrast, enhancement of respiration triggered cell death. CONCLUSION/SIGNIFICANCE: Thus, the Warburg effect might directly contribute to the initiation of cancer formation--not only by enhanced glycolysis--but also via decreased respiration in the presence of oxygen, which suppresses apoptosis.

Johnson D, Hilal N, Waters K, Hadler K, Cilliers J Langmuir. 2009 May 5;25(9):4880–5 We report the use of atomic force microscopy (AFM) to study the interactions between silica glass colloidal probes and charged microbubblescreated using one of two different surfactants: anionic sodium dodecyl sulfate (SDS) and cationic dodecyl trimethylammonium bromide (DTAB) in an aqueous environment. On close approach between the glass probe and a SDS microbubble, an appreciable repulsive force was observed prior to contact. This was not observed when using a DTAB microbubble, where only attractive forces were observed prior to contact. zeta-potential analysis showed that silica surfaces are negatively charged across the pH range of 3-10 when surfactant is not present. Addition of SDS did not alter the zeta-potential significantly, indicating that adsorption onto the particle surface did not occur. Conversely, the addition of DTAB decreased the negativity of the zeta-potential, reversing the sign, indicating that adsorption had occurred. This analysis was used in the removal of fine particlesfrom suspension using charged microbubbles. Silica particles were recovered using positively charged microbubbles from DTAB but not when using negatively charged microbubbles generated from SDS. Taken together, the data suggest that repulsive long-range interactions were responsible for the selective attachment of silica particles to microbubbles in a charge-dependent manner.

Hayward DC, Dosztányi Z, Clark-Walker GD PLoS ONE. 2013;8(2):e56465 The mitochondrial genome maintenance gene, MGM101, is essential for yeasts that depend on mitochondrial DNA replication. Previously, in Saccharomyces cerevisiae, it has been found that the carboxy-terminal two-thirds of Mgm101p has a functional core. Furthermore, there is a high level of amino acid sequence conservation in this region from widely diverse species. By contrast, the amino-terminal region, that is also essential for function, does not have recognizable conservation. Using a bioinformatic approach we find that the functional core from yeast and a corresponding region of Mgm101p from the coral Acropora millepora have an ordered structure, while the N-terminal domains of sequences from yeast and coral are predicted to be disordered. To examine whether ordered and disordered domains of Mgm101p have specific or general functions we made chimeric proteins from yeast and coral by swapping the two regions. We find, by an in vivo assay in S.cerevisiae, that the ordered domainof A.millepora can functionally replace the yeast core region but the disordered domain of the coral protein cannot substitute for its yeast counterpart.Mgm101p is found in the mitochondrial nucleoid along with enzymes and proteins involved in mtDNA replication. By attaching green fluorescent protein to the N-terminal disordered domain of yeast Mgm101p we find that GFP is still directed to the mitochondrial nucleoid where full-lengthMgm101p-GFP is targeted.

Choi D-H, Oh Y-M, Kwon S-H, Bae S-H J Microbiol. 2008 Feb;46(1):75–80 The SRL4 (YPL033C) gene was initially identified by the screening of Saccharomyces cerevisiae genes that play a role in DNA metabolism and/or genome stability using the SOS system of Escherichia coli. In this study, we found that the srl4Delta mutant cells were resistant to the chemicals that inhibit nucleotide metabolism and evidenced higher dNTP levels than were observed in the wild-type cells in the presence of hydroxyurea. The mutant cells also showed a significantly faster growth rate and higher dNTP levels at low temperature (16 degrees C) than were observed in the wild-type cells, whereas we detected no differences in the growth rate at 30 degrees C. Furthermore, srl4Delta was shown to suppress the lethality ofmutations of the essential S phase checkpoint genes, RAD53 and LCD1. These results indicate that SRL4 may be involved in the regulation of dNTPproduction by its function as a negative regulator of ribonucleotide reductase.

Naumov GI, Masneuf I, Naumova ES, Aigle M, Dubourdieu D Res Microbiol. 2000 Oct;151(8):683–91 Using genetic hybridization analysis, electrophoretic karyotyping and PCR-RFLP of the MET2 gene, we found that the yeast Saccharomycesbayanus var. uvarum is associated with certain types of wines produced in the Val de Loire, Sauternes, and Jurancon regions. The average frequency of appearance of this yeast in the three regions of France was 41, 7 and 77%, respectively. In contrast, we did not find S. bayanus var.uvarum in red wines produced in the Bordeaux area. The results of this study, as well as the findings already reported on Tokay (Slovakia), Muscat (Crimea, Ukraine) and Amarone (Italy) wines, lead us to consider that distribution of S. bayanus var. uvarum yeast is connected with low temperature climatic conditions and/or wine technologies in which must fermentation is at least partially carried out at low temperatures (10-15 degrees C).

Ohkura N, Yaguchi H, Tsukada T, Yamaguchi K Journal of Biological Chemistry. 2001 Oct 22;277(1):535–43 In extraskeletal myxoid chondrosarcoma, chromosomal translocation creates a gene fusion between EWS and the orphan nuclear receptor NOR1. The resulting fusion gene product, EWS/NOR1, has been believed to lead to malignant transformation by functioning as a transcriptional activator, but an alternative mechanism may also be involved. Here, using a newly developed functional complementation screening in yeast, we found thatEWS/NOR1, but not EWS or NOR1, complemented the loss of function of the small nuclear ribonucleoprotein Snu23p, an essential factor for pre-mRNA splicing in yeast. To verify the potential function of EWS/NOR1 in mammalian cells, we next showed that overexpression of EWS/NOR1caused increased usage of the distal 5'-splice site of pre-mRNA splicing and that EWS/NOR1 interacted with the human splicing protein U1C; neitherEWS nor NOR1 had the same activity or interaction as EWS/NOR1. Altogether, our findings reveal that EWS/NOR1 gains a novel activity affectingpre-mRNA splicing.

Warren DT, Andrews PD, Gourlay CW, Ayscough KR J Cell Sci. 2002 Apr 15;115(Pt 8):1703–15 Sla1p is a protein required for cortical actin patch structure and organisation in budding yeast. Here we use a combination of immunofluorescence microscopy and biochemical approaches to demonstrate interactions of Sla1p both with proteins regulating actin dynamics and with proteins required for endocytosis. Using Sla1p-binding studies we reveal association of Sla1p with two proteins known to be important for activation of the Arp2/3 complex in yeast, Abp1p and the yeast WASP homologue Las17p/Bee1p. A recent report of Sla1p association with Pan1p puts Sla1p in the currently unique position of being the only yeast protein known to interact with all three known Arp2/3-activating proteins in yeast. Localisation ofSla1p at the cell cortex is, however, dependent on the EH-domain-containing protein End3p, which is part of the yeast endocytic machinery. Using spectral variants of GFP on Sla1p (YFP) and on Abp1p (CFP) we show for the first time that these proteins can exist in discrete complexes at the cell cortex. However, the detection of a significant FRET signal means that these proteins also come close together in a single complex, and it is in this larger complex that we propose that Sla1p binding to Abp1p and Las17p/Bee1p is able to link actin dynamics to the endocytic machinery. Finally, we demonstrate marked defects in both fluid-phase and receptor-mediated endocytosis in cells that do not express SLA1, indicating thatSla1p is central to the requirement in yeast to couple endocytosis with the actin cytoskeleton.

Dewar H, Warren DT, Gardiner FC, Gourlay CG, Satish N, Richardson MR, Andrews PD, Ayscough KR Mol Biol Cell. 2002 Oct;13(10):3646–61 The importance of coupling the process of endocytosis to factors regulating actin dynamics has been clearly demonstrated in yeast, and manyproteins involved in these mechanisms have been identified and characterized. Here we demonstrate the importance of two additional cortical components, Ysc84p and Lsb5p, which together are essential for the organization of the actin cytoskeleton and for fluid phase endocytosis. Both Ysc84p and Lsb5p were identified through two-hybrid screens with different domains of the adaptor protein Sla1p. Ysc84p colocalizes with corticalactin and requires the presence of an intact actin cytoskeleton for its cortical localization. Ycl034w/Lsb5p localizes to the cell cortex but does not colocalize with actin. The Lsb5 protein contains putative VHS and GAT domains as well as an NPF motif, which are all domains characteristic ofproteins involved in membrane trafficking. Deletion of either gene alone does not confer any dramatic phenotype on cells. However, deletion of both genes is lethal at elevated temperatures. Furthermore, at all temperatures this double mutant has depolarized actin and an almost undetectable level of fluid phase endocytosis. Our data demonstrate that Ysc84p and Lsb5p are important components of complexes involved in overlapping pathways coupling endocytosis with the actin cytoskeleton in yeast.

Gourlay CW, Carpp LN, Timpson P, Winder SJ, Ayscough KR J Cell Biol. 2004 Mar 15;164(6):803–9 Several determinants of aging, including metabolic capacity and genetic stability, are recognized in both yeast and humans. However, many aspects of the pathways leading to cell death remain to be elucidated. Here we report a role for the actin cytoskeleton both in cell death and in promoting longevity. We have analyzed yeast strains expressing mutants with either increased or decreased actin dynamics. We show that decreased actindynamics causes depolarization of the mitochondrial membrane and an increase in reactive oxygen species (ROS) production, resulting in cell death. Important, however, is the demonstration that increasing actin dynamics, either by a specific actin allele or by deletion of a gene encoding the actin-bundling protein Scp1p, can increase lifespan by over 65%. Increased longevity appears to be due to these cells producing lower than wild-type levels of ROS. Homology between Scp1p and mammalian SM22/transgelin, which itself has been isolated in senescence screens, suggests a conserved mechanism linking aging to actin stability.

Marullo P, Bely M, Masneuf-Pomarede I, Aigle M, Dubourdieu D FEMS Yeast Res. 2004 May;4(7):711–9 Wine yeast strains exhibit a wide variability in their technological properties. The large number of allelic variants and the high degree of heterozygosity explain this genetic variability found among the yeast flora. Furthermore, most enological traits are controlled by polygenic systems presenting complex interactions between the alleles. Taking this into account, we hypothesized that the meiotic segregation of such alleles from a given strain might generate a progeny population with very different technological properties. In this work, a population of 50 progeny clones derived from four industrial wine strains of Saccharomyces cerevisiae was characterized for three major enological traits: ethanol tolerance, volatile-acidity production and hydrogen sulphide production. For this purpose, reliable laboratory fermentation tests were developed in accordance with enologicalpractice. A wide variability in the values of the various parameters was found among spore clones obtained after sporulation. Many clones presenting better aptitudes than the parental strains were obtained. Moreover, analysis of the progeny demonstrated that: (1) traits are in part inheritable; (2)traits are clearly polygenic; (3) broad relations of dominance/recessivity can be established. All these findings constitute an initial step for establishing breeding strategies for wine yeast improvement.