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235 records returned

Genetic evidence for a role of Saccharomyces cerevisiae Mph1 in recombinational DNA repair under replicative stressEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We describe a synthetic growth defect of an mph1 deletion if combined with an srs2 deletion that can result - depending on the genetic background - in synthetic lethality. The lethality is suppressed by mutations in homologous recombination (rad51, rad52, rad55, rad57) and in the DNA damage checkpoint (rad9, rad24, rad17). Importantly, rad54 and mph1, epistatic for damage sensitivity, are subadditive for spontaneous mutator phenotype. Therefore, Mph1 could be placed at the Rad51-mediated strand invasion process, with a function distinct from Rad54. Moreover, siz1 mutation is viable with mph1 and additive for DNA damage sen...
Source: Yeast - November 16, 2009 Category: Molecular Biology Authors: Evandro Rocco Panico, Christopher Ede, Michael Schildmann, Kirsten Anke Schürer, Wilfried Kramer Source Type: journals

The role of two putative nitroreductases, Frm2p and Hbn1p, in the oxidative stress response in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The nitroreductase family is comprised of a group of FMN- or FAD-dependent enzymes that are able to metabolize nitrosubstituted compounds using the reducing power of NAD(P)H. These nitroreductases can be found in bacterial species and, to a lesser extent, in eukaryotes. There is little information on the biochemical functions of nitroreductases. Some studies suggest their possible involvement in the oxidative stress response. In the yeast Saccharomyces cerevisiae, two nitroreductase proteins, Frm2p and Hbn1p, have been described. While Frm2p appears to act in the lipid signalling pathway, the function of Hbn1p is completel...
Source: Yeast - November 11, 2009 Category: Molecular Biology Authors: Iuri Marques de Oliveira, Alfeu Zanotto-Filho, José Cláudio Fonseca Moreira, Diego Bonatto, João Antonio Pêgas Henriques Source Type: journals

The human c-fos and TNF[alpha] AU-rich elements show different effects on mRNA abundance and protein expression depending on the reporter in the yeast Pichia pastorisEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In conclusion, we illustrate that the analysis of ARE-mediated effects on mRNA abundance and protein expression of a reporter depends on the sequence of the reporter itself as well as the ARE-surrounding sequences within the 3[prime] UTR. For this reason, we question whether already established reporter constructs in other cellular systems display the true type of regulation of the tested AREs for its original host gene. Finally, we propose that AREs should be analysed in their native sequence context. Copyright © 2009 John Wiley & Sons, Ltd. (Source: Yeast)
Source: Yeast - November 10, 2009 Category: Molecular Biology Authors: Thomas Lautz, Ulf Stahl, Christine Lang Source Type: journals

Paracoccin from Paracoccidioides brasiliensis; purification through affinity with chitin and identification of N-acetyl-[beta]-D-glucosaminidase activityEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The dimorphic fungus Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis, the most frequent systemic mycosis in Latin America. Our group has been working with paracoccin, a P. brasiliensis lectin with MM 70 kDa, which is purified by affinity with immobilized N-acetylglucosamine (GlcNAc). Paracoccin has been described to play a role in fungal adhesion to extracellular matrix components and to induce high and persistent levels of TNF[alpha] and nitric oxide production by macrophages. In the cell wall, paracoccin colocalizes with the [beta]-1,4-homopolymer of GlcNAc into the budding sites of the P. ...
Source: Yeast - November 10, 2009 Category: Molecular Biology Authors: Fausto Bruno dos Reis Almeida, Leandro Licursi de Oliveira, Marcelo Valle de Sousa, Maria Cristina Roque Barreira, Ebert Seixas Hanna Source Type: journals

Isolation and characterization of Candida kefyr orotidine-5[prime]-phosphate decarboxylase (URA3) geneEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Candida kefyr is a common yeast species that can be found in fermented milk and cheeses. As a first step to developing a gene transfer system for C. kefyr, the orotidine-5[prime]-phosphate decarboxylase (URA3) gene was cloned, using degenerate PCR and genome walking. The uninterrupted open reading frame of the C. kefyr URA3 gene spans 801 bp, corresponding to 267 amino acid residues. The functionality of the gene was confirmed by complementation of ura3 auxotrophs of C. albicans and Saccharomyces cerevisiae. Phylogenetic analysis of the deduced amino acid sequence indicated that it shares a high degree of homology with oth...
Source: Yeast - November 10, 2009 Category: Molecular Biology Authors: Paul Wai-Kei Tsang, Kin-Sing Wong, Jennifer Ka-Man Chu Source Type: journals

Dosage-dependent roles of the Cwt1 transcription factor for cell wall architecture, morphogenesis, drug sensitivity and virulence in Candida albicansEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The Cwt1 transcription factor is involved in cell wall architecture of the human fungal pathogen Candida albicans. We demonstrate here that deficiency of Cwt1 leads to decreased [beta]1,6-glucan in the cell wall, while mannoproteins are increased in the cell wall of exponentially growing cells and are released into the medium of stationary phase cells. Hyphal morphogenesis of cwt1 mutants is reduced on the surfaces of some inducing media. Unexpectedly, the CWT1/cwt1 heterozygous strains shows some stronger in vitro phenotypes compared to the homozygous mutant. The heterozygous but not the homozygous strain is also strongly...
Source: Yeast - November 10, 2009 Category: Molecular Biology Authors: Inmaculada Moreno, María Martinez-Esparza, Leslie Carolina Laforet, Rafael Sentandreu, Joachim F. Ernst, Eulogio Valentin Source Type: journals

Dual functions of Mdt1 in genome maintenance and cell integrity pathways in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Recent evidence indicates considerable cross-talk between genome maintenance and cell integrity control pathways. The RNA recognition motif (RRM)- and SQ/TQ cluster domain (SCD)-containing protein Mdt1 is required for repair of 3[prime]-blocked DNA double-strand breaks (DSBs) and efficient recombinational maintenance of telomeres in budding yeast. Here we show that deletion of MDT1 (PIN4/YBL051C) leads to severe synthetic sickness in the absence of the genes for the central cell integrity MAP kinases Bck1 and Slt2/Mpk1. Consistent with a cell integrity function, mdt1[Delta] cells are hypersensitive to the cell wall toxin c...
Source: Yeast - November 5, 2009 Category: Molecular Biology Authors: Ana Traven, Tricia L. Lo, Brietta L. Pike, Helena Friesen, Julie Guzzo, Brenda Andrews, Jörg Heierhorst Source Type: journals

Computational approaches for the genetic and phenotypic characterization of a Saccharomyces cerevisiae wine yeast collectionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Within this study, we have used a set of computational techniques to relate the genotypes and phenotypes of natural populations of Saccharomyces cerevisiae, using allelic information from 11 microsatellite loci and results from 24 phenotypic tests. A group of 103 strains was obtained from a larger S. cerevisiae winemaking strain collection by clustering with self-organizing maps. These strains were further characterized regarding their allelic combinations for 11 microsatellites and analysed in phenotypic screens that included taxonomic criteria (carbon and nitrogen assimilation tests, growth at different temperatures) and...
Source: Yeast - November 4, 2009 Category: Molecular Biology Authors: R. Franco-Duarte, L. Umek, B. Zupan, D. Schuller Source Type: journals

Random and targeted gene integrations through the control of non-homologous end joining in the yeast Kluyveromyces marxianusEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Kluyveromyces marxianus DMKU3-1042 is a thermotolerant yeast strain suitable for high-temperature ethanol fermentation and genetic engineering with linear DNA. We have developed a highly efficient random gene integration method with a frequency that exceeds 2.5 × 106 transformants/µg linear DNA, a figure comparable to what is observed with autonomously replicating plasmid transformation in Saccharomyces cerevisiae. To establish the mechanism of random integration in DMKU3-1042, we identified and deleted the K. marxianus KU70 gene, which is known to be involved in the non-homologous end-joining (NHEJ) pathway. In yeast la...
Source: Yeast - November 4, 2009 Category: Molecular Biology Authors: Babiker M. A. Abdel-Banat, Sanom Nonklang, Hisashi Hoshida, Rinji Akada Source Type: journals

Cellular and transcriptional responses of yeast to the cleavage of cytosolic tRNAs induced by colicin DEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Colicin D is a plasmid-encoded antibacterial protein that specifically cleaves the anticodon loops of four Escherichia coli tRNAArg species. Here, we report that the catalytic domain of colicin D, which is expressed in Saccharomyces cerevisiae, impairs cell growth by cleaving specific tRNAs. DNA microarray analysis revealed that mating-related genes were upregulated, while genes involved in a range of metabolic processes were downregulated, thereby impairing cell growth. The pheromone-signalling pathway was activated only in [alpha] cells by tRNA cleavage, which was not observed in 'a' cells or diploid cells. On the basis ...
Source: Yeast - October 29, 2009 Category: Molecular Biology Authors: Megumi Shigematsu, Tetsuhiro Ogawa, Atsuhiro Kido, Hiroko K. Kitamoto, Makoto Hidaka, Haruhiko Masaki Source Type: journals

RRD1, a component of the TORC1 signalling pathway, affects anaesthetic response in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The molecular mechanisms of action of volatile anaesthetics remain unknown despite clinical use for over 150 years. While many effects of these agents have been characterized, clear insight into how these effects relate to the physiological state of anaesthesia has not been established. Volatile anaesthetics arrest cell division in Saccharomyces cerevisiae in a manner that parallels the anaesthetic actions of these drugs in mammals. To gain additional insight into the cellular activities of these drugs, we isolated genes that, when present on multi-copy plasmids, render S. cerevisiae resistant to the volatile anaesthetic i...
Source: Yeast - September 22, 2009 Category: Molecular Biology Authors: Laura K. Palmer, Beverly A. Baptiste, John C. Fester, Justin C. Perkins, Ralph L. Keil Source Type: journals

Prion-associated proteins in yeast: comparative analysis of isogenic [PSI+] and [psi-] strainsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
A large group of prion-associated proteins was identified in yeast cells using a new approach, comparative analysis of pellet proteins of crude cell lysates in isogenic strains of Saccharomyces cerevisiae differing by their prion composition. Two-dimensional (2D) electrophoresis followed by MALDI analysis of the pellet proteins of [PSI+] and [psi-] strains after prion elimination by GuHCl and prion transmission by cytoduction permitted identification of ca. 40 proteins whose aggregation state correlated with the change of prion(s) content. Approximately half of these proteins belonged to chaperones and to enzymes of glucos...
Source: Yeast - September 21, 2009 Category: Molecular Biology Authors: Olga V. Nevzglyadova, Alexey V. Artemov, Alexey G. Mittenberg, Kirill V. Solovyov, Elena I. Kostyleva, Ekaterina V. Mikhailova, Irina M. Kuznetsova, Konstantin K. Turoverov, Tonu R. Soidla Source Type: journals

Yap4 PKA- and GSK3-dependent phosphorylation affects its stability but not its nuclear localizationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Yap4 is a nuclear-resident transcription factor induced in Saccharomyces cerevisiae when exposed to several stress conditions, which include mild hyperosmotic and oxidative stress, temperature shift or metal exposure. This protein is also phosphorylated. Here we report that this modification is driven by PKA and GSK3. In order to ascertain whether Yap4 is directly or indirectly phosphorylated by PKA, we searched for stress and PKA-related kinases that could phosphorylate Yap4. We show that phosphorylation is independent of the kinases Rim15, Yak1, Sch9, Slt2, Ste20 and Ptk2. In addition, we showed that Yap4 phosphorylation...
Source: Yeast - September 21, 2009 Category: Molecular Biology Authors: Jorge Pereira, Catarina Pimentel, Catarina Amaral, Regina A. Menezes, Claudina Rodrigues-Pousada Source Type: journals

Yeast Ste23p shares functional similarities with mammalian insulin-degrading enzymesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, recombinant Ste23p and R. norvegicus IDE (RnIDE) were isolated from E. coli, and their enzymatic properties compared. Ste23p was found to cleave established RnIDE substrates, including the amyloid-[beta] peptide (A[beta]1-40) and insulin B-chain. A novel internally quenched fluorogenic substrate (Abz-SEKKDNYIIKGV-nitroY-OH) based on the polypeptide sequence of the yeast P2 a-factor mating propheromone was determined to be a suitable substrate for both Ste23p and RnIDE, and was used to conduct comparative enzymological studies. Both enzymes were most active at 37 °C, in alkaline buffers and in high salt envi...
Source: Yeast - September 11, 2009 Category: Molecular Biology Authors: Benjamin J. Alper, Jarrad W. Rowse, Walter K. Schmidt Source Type: journals

MPR1 as a novel selection marker in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
L-Azetidine-2-carboxylic acid (AZC) is a toxic four-membered ring analogue of L-proline that is transported into cells by proline transporters. AZC and L-proline in the cells are competitively incorporated into nascent proteins. When AZC is present in a minimum medium, misfolded proteins are synthesized in the cells, thereby inhibiting cell growth. The MPR1 gene has been isolated from the budding yeast Saccharomyces cerevisiae [Sigma]1278b as a multicopy suppressor of AZC-induced growth inhibition. MPR1 encodes a novel acetyltransferase that detoxifies AZC via N-acetylation. Since MPR1 is absent in the laboratory strain of...
Source: Yeast - September 10, 2009 Category: Molecular Biology Authors: Kaoru Ogawa-Mitsuhashi, Koji Sagane, Junro Kuromitsu, Hiroshi Takagi, Kappei Tsukahara Source Type: journals

Microarray studies on the genes responsive to the addition of spermidine or spermine to a Saccharomyces cerevisiae spermidine synthase mutantEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The naturally occurring polyamines putrescine, spermidine or spermine are ubiquitous in all cells. Although polyamines have prominent regulatory roles in cell division and growth, precise molecular and cellular functions are not well-established in vivo. In this work we have performed microarray experiments with a spermidine synthase, spermine oxidase mutant ([Delta]spe3 [Delta]fms1) strain to investigate the responsiveness of yeast genes to supplementation with spermidine or spermine. Expression analysis identified genes responsive to the addition of either excess spermidine (10-5 M) or spermine (10-5 M) compared to a con...
Source: Yeast - August 16, 2009 Category: Molecular Biology Authors: Manas K. Chattopadhyay, Weiping Chen, George Poy, Margaret Cam, David Stiles, Herbert Tabor Source Type: journals

Characterization of chromosomal integration sites for heterologous gene expression in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The construction of mitotically stable yeast strains for heterologous gene or pathway expression often requires chromosomal integration. However, transcription levels vary between different chromosome regions. We therefore characterized 20 different integration sites of the Sacchromyces cerevisiae genome by inserting lacZ as a reporter gene under the control of two different promoters and determining expression levels through enzyme activity measurement. An up to 8.7-fold difference was detected between the sites conferring lowest and highest expression, respectively. This opens the opportunity for modulating gene expressi...
Source: Yeast - August 13, 2009 Category: Molecular Biology Authors: Dongmei Bai Flagfeldt, Verena Siewers, Le Huang, Jens Nielsen Source Type: journals

In memoriam: Piotr Slonimski (9 November 1922-25 April 2009)Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
No Abstract. (Source: Yeast)
Source: Yeast - August 12, 2009 Category: Molecular Biology Authors: Christopher J. Herbert Source Type: journals

Efficient production of L-lactic acid by Crabtree-negative yeast Candida boidiniiEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Industrial production of L-lactic acid, which in polymerized form as poly-lactic acid is widely used as a biodegradable plastic, has been attracting world-wide attention. By genetic engineering we constructed a strain of the Crabtree-negative yeast Candida boidinii that efficiently produced a large amount of L-lactic acid. The alcohol fermentation pathway of C. boidinii was altered by disruption of the PDC1 gene encoding pyruvate decarboxylase, resulting in an ethanol production that was reduced to 17% of the wild-type strain. The alcohol fermentation pathway of the PDC1 deletion strain was then successfully utilized for t...
Source: Yeast - August 3, 2009 Category: Molecular Biology Authors: Fumi Osawa, Toshio Fujii, Takehisa Nishida, Nobuki Tada, Toru Ohnishi, Osamu Kobayashi, Toshihiro Komeda, Satoshi Yoshida Source Type: journals

New selectable host-marker systems for multiple genetic manipulations based on TRP1, MET2 and ADE2 in the methylotrophic yeast Hansenula polymorphaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Interest has been increasing in the thermotolerant methylotrophic yeast Hansenula polymorpha as a useful system for fundamental research and applied purposes. Only a few genetic marker genes and auxotrophic hosts are yet available for this yeast. Here we isolated and developed H. polymorpha TRP1, MET2 and ADE2 genes as selectable markers for multiple genetic manipulations. The H. polymorpha TRP1 (HpTRP1), MET2 (HpMET2) and ADE2 (HpADE2) genes were sequentially disrupted, using an HpURA3 pop-out cassette in H. polymorpha to generate a series of new multiple auxotrophic strains, including up to a quintuple auxotrophic strain...
Source: Yeast - August 2, 2009 Category: Molecular Biology Authors: Seon Ah Cheon, Jinho Choo, Vera M. Ubiyvovk, Jeong-Nam Park, Moo Woong Kim, Doo-Byoung Oh, Ohsuk Kwon, Andriy A. Sibirny, Jeong-Yoon Kim, Hyun Ah Kang Source Type: journals

Improved gap-repair cloning method that uses oligonucleotides to target cognate sequencesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In vivo or gap-repair cloning in yeast has been widely recognized as one of the most efficient means for error-free construction of plasmids. A protocol is described here that allows easy and efficient gap-repair cloning that is based on two major modifications. Instead of subcloning, the targeting plasmids are constructed using oligonucleotides from sequences derived from the upstream and downstream sequences of the fragment to be cloned. These sequences are selected so that they can lead to the generation of recognition sites for restriction enzymes that produce blunt ends. Accordingly, this procedure can be applied to a...
Source: Yeast - July 20, 2009 Category: Molecular Biology Authors: Ana A. Kitazono Source Type: journals

Selection of cell death-deficient p53 mutants in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study we explored the effect of other p53 mutants, such as the hot-spot mutant R282W and the double mutant N268S::I332V. Unexpectedly, both mutants behaved inversely to R248W, as they completely inhibited yeast growth on minimal medium and induced ROS production. This phenotype 'yeast cell death on minimal medium' allowed for the subsequent screening of intragenic p53-inactivating mutations. In all cases, the 'revertant yeast clones' display a complete p53 inactivation through either gross deletion or nonsense mutations. More interestingly, missense mutations were also found: the deletion of I255 or substitution of...
Source: Yeast - July 4, 2009 Category: Molecular Biology Authors: Ines Yacoubi-Hadj Amor, Kamel Smaoui, Hanène Belguith, Lamia Djemal, Mosbeh Dardouri, Raja Mokdad-Gargouri, Ali Gargouri Source Type: journals

Fed-batch methanol feeding strategy for recombinant protein production by Pichia pastoris in the presence of co-substrate sorbitolEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Batch-wise sorbitol addition as a co-substrate at the induction phase of methanol fed-batch fermentation by Pichia pastoris (Mut+) was proposed as a beneficial recombinant protein production strategy and the metabolic responses to methanol feeding rate in the presence of sorbitol was systematically investigated. Adding sorbitol batch-wise to the medium provided the following advantages over growth on methanol alone: (a) eliminating the long lag-phase for the cells and reaching 'high cell density production' at t = 24 h of the process (CX = 70 g CDW/l); (b) achieving 1.8-fold higher recombinant human erythropoietin (rHuEPO)...
Source: Yeast - July 2, 2009 Category: Molecular Biology Authors: Eda Çelik, P[inodot]nar Çal[inodot]k, Stephen G. Oliver Source Type: journals

Identification of proteins associated with the yeast mitochondrial RNA polymerase by tandem affinity purificationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The abundance of mitochondrial (mt) transcripts varies under different conditions, and is thought to depend upon rates of transcription initiation, transcription termination/attenuation and RNA processing/degradation. The requirement to maintain the balance between RNA synthesis and processing may involve coordination between these processes; however, little is known about factors that regulate the activity of mtRNA polymerase (mtRNAP). Recent attempts to identify mtRNAP-protein interactions in yeast by means of a generalized tandem affinity purification (TAP) protocol were not successful, most likely because they involved...
Source: Yeast - June 24, 2009 Category: Molecular Biology Authors: Dmitriy A. Markov, Maria Savkina, Michael Anikin, Mark Del Campo, Karen Ecker, Alan M. Lambowitz, Jon P. De Gnore, William T. McAllister Source Type: journals

A QPCR-based reporter system to study post-transcriptional regulation via the 3[prime] untranslated region of mRNA in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Post-transcriptional regulation via the 3[prime] untranslated region (3[prime] UTR) of mRNA is an important factor in governing eukaryotic gene expression. Achieving detailed understanding of these processes requires highly quantitative systems in which comparative studies can be performed. To this end, we have developed a plasmid reporter system for Saccharomyces cerevisiae, in which the 3[prime] UTR can be easily replaced and modified. Accurate quantification of the tandem affinity purification tag (TAP)-reporter protein and of TAP-mRNA is achieved by immuno-QPCR and by RT-QPCR, respectively. We have used our reporter sy...
Source: Yeast - June 8, 2009 Category: Molecular Biology Authors: Kristina Lind, Joakim Norbeck Source Type: journals

Baker's yeast expressing the Japanese encephalitis virus envelope protein on its cell surface: induction of an antigen-specific but non-neutralizing antibody responseEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Live recombinant Saccharomyces cerevisiae yeast expressing the envelope antigen of Japanese encephalitis virus (JEV) on the outer mannoprotein layer of the cell wall were examined for their ability to induce antigen-specific antibody responses in mice. When used as a model antigen, parenteral immunization of mice with surface-expressing GFP yeast induced a strong anti-GFP antibody response in the absence of adjuvants. This antigen delivery approach was then used for a more stringent system, such as the envelope protein of JEV, which is a neurotropic virus requiring neutralizing antibodies for protection. Although 70% of ce...
Source: Yeast - June 8, 2009 Category: Molecular Biology Authors: Bhaskar Upadhyaya, Ramanathapuram Manjunath Source Type: journals

Proteolytic processing of certain CaaX motifs can occur in the absence of the Rce1p and Ste24p CaaX proteasesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The CaaX motif directs C-terminal protein modifications that include isoprenylation, proteolysis and carboxylmethylation. Proteolysis is generally believed to require either Rce1p or Ste24p. While investigating the substrate specificity of these proteases, using the yeast a-factor mating pheromone as a reporter, we observed Rce1p- and Ste24p-independent mating (RSM) when the CKQQ CaaX motif was used in lieu of the natural a-factor CVIA motif. Uncharged or negatively charged amino acid substitutions at the a1 position of the CKQQ motif prevented RSM. Alanine substitutions at the a2 and X positions enhanced RSM. Random mutag...
Source: Yeast - June 7, 2009 Category: Molecular Biology Authors: Ranjith K. Krishnankutty, Sayali S. Kukday, Amanda J. Castleberry, Sarah R. Breevoort, Walter K. Schmidt Source Type: journals

Proteome analysis of the xylose-fermenting mutant yeast strain TMB 3400Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, the mutant S. cerevisiae strain TMB 3400, which has good xylose fermentation properties, was compared with its parental strain to examine the factors behind the improved xylose utilization at protein level. The proteome of the parental and the mutant strains were characterized by difference in gel electrophoresis (DiGE) to quantitatively identify proteins that are expressed at altered levels in the mutant. The most significant changes detected by proteome analysis were the 6-10-fold increased levels of xylose reductase, xylitol dehydrogenase and transketolase (Tkl1) in the mutant, which is in accordance with...
Source: Yeast - June 7, 2009 Category: Molecular Biology Authors: Kaisa Karhumaa, Anna-Karin Påhlman, Bärbel Hahn-Hägerdal, Fredrik Levander, Marie-F. Gorwa-Grauslund Source Type: journals

Additional cassettes for epitope and fluorescent fusion proteins in Candida albicansEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Epitope tags that confer specific properties, including affinity for resins or antibodies or detection by fluorescence microscopy, are highly useful for biochemical and cell biological investigations. In Candida albicans and several other related yeasts, the CUG codon specifies serine instead of leucine, requiring that molecular tools be customized for use in this important human fungal pathogen. Here we report the construction of a set of plasmids containing 13-Myc, 3HA, GST, V5 or His9 epitope cassettes that facilitate PCR-mediated construction of epitope-tagged proteins. Common primer sets amplify the different tags wit...
Source: Yeast - June 7, 2009 Category: Molecular Biology Authors: Maryam Gerami-Nejad, Keely Dulmage, Judith Berman Source Type: journals

Quantitation of NAD+ biosynthesis from the salvage pathway in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Nicotinamide adenine dinucleotide (NAD+) is synthesized via two major pathways in prokaryotic and eukaryotic systems: the de novo biosynthesis pathway from tryptophan precursors, or the salvage biosynthesis pathway from either extracellular nicotinic acid or various intracellular NAD+ decomposition products. NAD+ biosynthesis via the salvage pathway has been linked to an increase in yeast replicative lifespan under calorie restriction (CR). However, the relative contribution of each pathway to NAD+ biosynthesis under both normal and CR conditions is not known. Here, we have performed lifespan, NAD+ and NADH (the reduced fo...
Source: Yeast - April 29, 2009 Category: Molecular Biology Authors: Jennifer Sporty, Su-Ju Lin, Michiko Kato, Ted Ognibene, Benjamin Stewart, Ken Turteltaub, Graham Bench Source Type: journals

Expression of GFP using Pichia pastoris vectors with zeocin or G-418 sulphate as the primary selectable markerEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Pichia pastoris is a popular host organism for expressing heterologous proteins, and various expression vectors for this yeast are currently available. Recently, vectors containing novel dominant antibiotic resistance markers have become a strong and developing field of research for this methylotropic yeast strain. We have developed new P. pastoris expression vectors, the pPICKanMX6 and pPICKanMX6[alpha] series. These vectors were constructed by replacing the zeocin resistance gene of the pPICZA, B, C and pPICZ[alpha]A, B and C vectors with the Tn903 kanR marker from pFA6a KanMX6, which confers G-418 sulphate resistance in...
Source: Yeast - April 27, 2009 Category: Molecular Biology Authors: Theo Papakonstantinou, Simon Harris, Milton T. W. Hearn Source Type: journals

The Hsp90/Cdc37p chaperone system is a determinant of molybdate resistance in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Saccharomyces cerevisiae lacks enzymes that contain the molybdopterin co-factor and therefore any requirement for molybdenum as a trace mineral supplement. Instead, high molybdate levels are inhibitory to its growth. Low cellular levels of heat shock protein 90 (Hsp90), an essential chaperone, were found to enhance this sensitivity to molybdate. Certain Hsp90 point mutations and co-chaperone protein defects that partially compromise the function of the Hsp90/Cdc37p chaperone system also rendered S. cerevisiae hypersensitive to high molybdate levels. Sensitivity was especially apparent with mutations close to the Hsp90 nucl...
Source: Yeast - April 27, 2009 Category: Molecular Biology Authors: Stefan H. Millson, James M. Nuttall, Mehdi Mollapour, Peter W. Piper Source Type: journals

Atan1p - an extracellular tannase from the dimorphic yeast Arxula adeninivorans: molecular cloning of the ATAN1 gene and characterization of the recombinant enzymeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The tannase-encoding Arxula adeninivorans gene ATAN1 was isolated from genomic DNA by PCR, using as primers oligonucleotide sequences derived from peptides obtained after tryptic digestion of the purified tannase protein. The gene harbours an ORF of 1764 bp, encoding a 587-amino acid protein, preceded by an N-terminal secretion sequence comprising 28 residues. The deduced amino acid sequence was similar to those of tannases from Aspergillus oryzae (50% identity), A. niger (48%) and putative tannases from A. fumigatus (52%) and A. nidulans (50%). The sequence contains the consensus pentapeptide motif (-Gly-X-Ser-X-Gly-) whi...
Source: Yeast - April 22, 2009 Category: Molecular Biology Authors: Erik Böer, Rüdiger Bode, Hans-Peter Mock, Michael Piontek, Gotthard Kunze Source Type: journals

Sdo1p, the yeast orthologue of Shwachman-Bodian-Diamond syndrome protein, binds RNA and interacts with nuclear rRNA-processing factorsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The Shwachman-Bodian-Diamond syndrome protein (SBDS) is a member of a highly conserved protein family of not well understood function, with putative orthologues found in different organisms ranging from Archaea, yeast and plants to vertebrate animals. The yeast orthologue of SBDS, Sdo1p, has been previously identified in association with the 60S ribosomal subunit and is proposed to participate in ribosomal recycling. Here we show that Sdo1p interacts with nucleolar rRNA processing factors and ribosomal proteins, indicating that it might bind the pre-60S complex and remain associated with it during processing and transport ...
Source: Yeast - April 18, 2009 Category: Molecular Biology Authors: Juliana S. Luz, Raphaela C. Georg, Carlos H. Gomes, Gláucia M. Machado-Santelli, Carla C. Oliveira Source Type: journals

Sequence of the yeast protein expression plasmid pEG(KT)Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The plasmid pEG(KT) is a widely used plasmid for expressing high levels of GST fusion proteins in the yeast Saccharomyces cerevisiae. Unfortunately, a complete sequence file has been lacking, thus complicating efforts to design cloning projects or to modify the plasmid for other uses (e.g. exchanging selection markers, epitope tags or protease cleavage sites to remove the epitope tag). Here, the complete sequence of the pEG(KT) plasmid is reported, thus facilitating its use. Additionally, its use as a vector backbone for high-level expression of a TAP-tagged protein is shown. Copyright © 2009 John Wiley & Sons, Ltd. (Source: Yeast)
Source: Yeast - April 6, 2009 Category: Molecular Biology Authors: B. Daniel Pierce, Beverly Wendland Source Type: journals

Elizabeth W. Jones, 8 March 1939-11 June 2008Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
No Abstract. (Source: Yeast)
Source: Yeast - March 29, 2009 Category: Molecular Biology Authors: Carol S. Newlon Source Type: journals

The mating response cascade does not modulate changes in the steady-state level of target mRNAs through control of mRNA stabilityEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Many extracellular signals trigger changes in gene expression by altering the steady-state level of target transcripts. This modulation of transcript levels is typically ascribed to changes in transcription of target genes; however, there are numerous examples of changes in mRNA processing and stability that contribute to the overall change in transcript levels following signalling pathway activation. The [alpha]-factor-stimulated mating pathway in Saccharomyces cerevisiae is a receptor-operated MAP kinase cascade that results in increased levels of a large number of target mRNA transcripts when stimulated acutely. A previ...
Source: Yeast - March 24, 2009 Category: Molecular Biology Authors: Chad M. Kitchen, Sara W. Leung, Anita H. Corbett, T. J. Murphy Source Type: journals

Expression profiling of the bottom fermenting yeast Saccharomyces pastorianus orthologous genes using oligonucleotide microarraysEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The bottom fermenting yeast Saccharomyces pastorianus is reported to have arisen as a natural hybrid of two yeast strains, S. cerevisiae and S. bayanus. The S. pastorianus genome includes S. cerevisiae-type (Sc-type) genes and orthologous lager-fermenting-yeast specific-type (Lg-type) genes derived from S. cerevisiae and S. bayanus, respectively. To gain insights into the physiological properties of S. pastorianus, we developed an in situ synthesized 60-mer oligonucleotide microarray for gene expression monitoring of these orthologous genes, consisting of approximately 6600 Sc-type genes and 3200 Lg-type genes. A compariso...
Source: Yeast - February 26, 2009 Category: Molecular Biology Authors: Toshiko Minato, Satoshi Yoshida, Tatsuji Ishiguro, Emiko Shimada, Satoru Mizutani, Osamu Kobayashi, Hiroyuki Yoshimoto Tags: Research Articles Source Type: journals

Current awareness on yeastEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In order to keep subscribers up-to-date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly-published material on yeasts. Each bibliography is divided into 10 sections. 1 Reviews; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (8 we...
Source: Yeast - February 25, 2009 Category: Molecular Biology Authors: John Wiley & Sons, Ltd. Tags: Current Awareness Source Type: journals

Small epitope-linker modules for PCR-based C-terminal tagging in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
PCR-mediated gene modification is a powerful approach to the functional analysis of genes in Saccharomyces cerevisiae. One application of this method is epitope-tagging of a gene to analyse the corresponding protein by immunological methods. However, the number of epitope tags available in a convenient format is still low, and interference with protein function by the epitope, particularly if it is large, is not uncommon. To address these limitations and broaden the utility of the method, we constructed a set of convenient template plasmids designed for PCR-based C-terminal tagging with 10 different, relatively short pepti...
Source: Yeast - February 25, 2009 Category: Molecular Biology Authors: Minoru Funakoshi, Mark Hochstrasser Tags: Research Articles Source Type: journals

Production of polyunsaturated fatty acids in yeast Saccharomyces cerevisiae and its relation to alkaline pH toleranceEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Saccharomyces cerevisiae produces saturated and monounsaturated fatty acids of 16- and 18-carbon atoms and no polyunsaturated fatty acids (PUFAs) with more than two double bonds. To study the biological significance of PUFAs in yeast, we introduced Kluyveromyces lactis [Delta]12 fatty acid desaturase (KlFAD2) and [omega]3 fatty acid desaturase (KlFAD3) genes into S. cerevisiae to produce linoleic and [alpha]-linolenic acids in S. cerevisiae. The strain producing linoleic and [alpha]-linolenic acids showed an alkaline pH-tolerant phenotype. DNA microarray analyses showed that the transcription of a set of genes whose expres...
Source: Yeast - February 25, 2009 Category: Molecular Biology Authors: Hisashi Yazawa, Hitoshi Iwahashi, Yasushi Kamisaka, Kazuyoshi Kimura, Hiroshi Uemura Tags: Research Articles Source Type: journals

Characterization and expression analysis of a gene cluster for nitrate assimilation from the yeast Arxula adeninivoransEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In Arxula adeninivorans nitrate assimilation is mediated by the combined actions of a nitrate transporter, a nitrate reductase and a nitrite reductase. Single-copy genes for these activities (AYNT1, AYNR1, AYNI1, respectively) form a 9103 bp gene cluster localized on chromosome 2. The 3210 bp AYNI1 ORF codes for a protein of 1070 amino acids, which exhibits a high degree of identity to nitrite reductases from the yeasts Pichia anomala (58%), Hansenula polymorpha (58%) and Dekkera bruxellensis (54%). The second ORF (AYNR1, 2535 bp) encodes a nitrate reductase of 845 residues that shows significant (51%) identity to nitrate ...
Source: Yeast - February 5, 2009 Category: Molecular Biology Authors: Erik Böer, Anja Schröter, Rüdiger Bode, Michael Piontek, Gotthard Kunze Source Type: journals

Current awareness on yeastEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In order to keep subscribers up-to-date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly-published material on yeasts. Each bibliography is divided into 10 sections. 1 Reviews; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (5 we...
Source: Yeast - January 29, 2009 Category: Molecular Biology Authors: John Wiley & Sons, Ltd. Tags: Current Awareness Source Type: journals

SpOPT1, a member of the oligopeptide family (OPT) of the fission yeast Schizosaccharomyces pombe, is involved in the transport of glutathione through the outer membrane of the cellEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
A protein involved in the transport of glutathione has been identified, cloned and characterized from the fission yeast Schizosaccharomyces pombe. Database searches revealed the Sz. pombe ORF SPAC29B12.10c as a close homologue to several members of the OPT family, including the Saccharomyces cerevisiae high-affinity glutathione transporter Hgt1p. The gene product of SPAC29B12.10c has been identified as a protein, named SpOPT1, localized within the plasma membrane, transporting the tripeptide glutathione. Disruption of SPAC29B12.10c led to strains inable to grow on media containing glutathione as a sole source of sulphur, d...
Source: Yeast - January 29, 2009 Category: Molecular Biology Authors: Tamara Dworeck, Klaus Wolf, Martin Zimmermann Tags: Yeast Functional Analysis Reports Source Type: journals

Plasmids with E2 epitope tags: tagging modules for N- and C-terminal PCR-based gene targeting in both budding and fission yeast, and inducible expression vectors for fission yeastEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This report presents a series of plasmids for the E2 epitope tagging of proteins in Saccharomyces cerevisiae and Schizosaccharomyces pombe. E2Tags are 10-amino acids (epitope E2a: SSTSSDFRDR)- and 12 amino acids (epitope E2b: GVSSTSSDFRDR)-long peptides derived from the E2 protein of bovine papillomavirus type 1. The modules for C-terminal tagging with E2a and E2b epitopes were constructed by the modification of the pYM-series plasmid. The N-terminal E2a and E2b tagging modules were based on pOM-series plasmid. The pOM-series plasmids were selected for this study because of their use of the Cre-loxP recombination system. T...
Source: Yeast - January 29, 2009 Category: Molecular Biology Authors: Tiina Tamm Tags: Yeast Functional Analysis Reports Source Type: journals

HIV-1 integrase trafficking in S. cerevisiae: a useful model to dissect the microtubule network involvement of viral protein nuclear importEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Intracellular transport of karyophilic cargos comprises translocation to the nuclear envelope and subsequent nuclear import. Small cargos such as isolated proteins can reach the nuclear envelope by diffusion but movement of larger structures depends on active translocation, typically using microtubules. Centripetal transport ends at the perinuclear microtubule organizing centre called the spindle pole body (SPB) in yeast. Previously, we found by two hybrids that the karyophilic lentiviral-encoded integrase (IN) interacts with two yeast microtubule-associated proteins, Dyn2p (dynein light chain protein) and Stu2p, a centros...
Source: Yeast - January 29, 2009 Category: Molecular Biology Authors: S. Desfarges, B. Salin, C. Calmels, M. L. Andreola, V. Parissi, M. Fournier Tags: Research Articles Source Type: journals

PGK1, the gene encoding the glycolitic enzyme phosphoglycerate kinase, acts as a multicopy suppressor of apoptotic phenotypes in S. cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In a previous paper we reported the construction of a S. cerevisiae strain lacking the essential gene LSM4, which could survive by the introduction of a truncated form of the orthologous gene from Kluyveromyces lactis. This strain showed apoptotic hallmarks and other phenotypes, including an increased sensitivity to caffeine and acetic acid. The suppression of the latter phenotype by overexpressing yeast genes allowed the isolation of PGK1, the gene encoding the glycolytic enzyme phosphoglycerate kinase. This gene restored normal ageing, oxygen peroxide resistance and nuclear integrity in the mutant. Other phenotypes, such...
Source: Yeast - January 29, 2009 Category: Molecular Biology Authors: Cristina Mazzoni, Mirko Torella, Agnese Petrera, Vanessa Palermo, Claudio Falcone Tags: Research Articles Source Type: journals

Effect of trehalose accumulation on response to saline stress in Saccharomyces cerevisiaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
To examine the effect of trehalose accumulation on response to saline stress in Saccharomyces cerevisiae, we constructed deletion strains of all combinations of the trehalase genes ATH1, NTH1 and NTH2 and examined their growth behaviour and intracellular trehalose accumulation under non-stress and saline-stress conditions. Saline stress was induced in yeast cells by NaCl addition at the exponential growth phase. All deletion strains showed similar specific growth rates and trehalose accumulation to their parent strain under non-stress conditions. However, under the saline stress condition, one single deletion strain, nth1[...
Source: Yeast - January 29, 2009 Category: Molecular Biology Authors: Siraje Arif Mahmud, Keisuke Nagahisa, Takashi Hirasawa, Katsunori Yoshikawa, Kengo Ashitani, Hiroshi Shimizu Tags: Research Articles Source Type: journals

Current awareness on yeastEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In order to keep subscribers up-to-date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly-published material on yeasts. Each bibliography is divided into 10 sections. 1 Reviews; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (5 we...
Source: Yeast - January 21, 2009 Category: Molecular Biology Authors: John Wiley & Sons, Ltd. Tags: Current Awareness Source Type: journals

Joined in death: highlights of the Sixth International Meeting on Yeast Apoptosis in Leuven, Belgium, 30 April-4 May 2008Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
While yeast apoptosis was still a controversial issue less than 10 years ago, the efforts of many groups have revealed cell death mechanisms that resemble, in many aspects, those described for mammalian apoptosis. Here, we provide an overview of new insights on yeast apoptosis and the link with lifespan of yeast cells, based on data presented at the 6th International Meeting of Yeast Apoptosis (IMYA). Together, these data demonstrate the power and advantages of the yeast system to uncover novel cellular factors governing life and death, placing yeast at the forefront of apoptosis research. Copyright © 2009 John Wiley & So...
Source: Yeast - January 21, 2009 Category: Molecular Biology Authors: Karin Thevissen, Frank Madeo, Paula Ludovico, Bruno Cammue, Joris Winderickx Tags: Conference Reports Source Type: journals