bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of plasma membrane proton‐ATPase from salt‐tolerant yeast Zygosaccharomyces rouxii cells
Authors: Watanabe Yasuo , Yamaguchi Masahiro , Sanemitsu Yumi , Tamai Youichi 
Published: May 2005

AbstractPlasma membrane was isolated from the salt‐tolerant yeast Zygosaccharomyces rouxii and from Saccharomyces cerevisiae. The ATPase in the plasma membrane of Z. rouxii cells was a typical proton‐ATPase as judged by testing with various ATPase inhibitors. There were slight differences in the pH optima of activities and in the sensitivity to sodium chloride (NaCl) and potassium chloride (KCl) of the ATPase from Z. rouxii and S. cerevisiae. The specific ATPase activity from Z. rouxii was higher in cells grown in a medium containing 2 M‐NaCl than in those not containing NaCl. No in vivo activation by incubation with glucose was observed in Z. rouxii cells and the specific ATPase activity was independent of the growth phase, unlike S. cerevisiae cells.
Intracellular expression of Kluyveromyces lactis toxin γ subunit mimics treatment with exogenous toxin and distinguishes two classes of toxin‐resistant mutant
Authors: Butler Andrew R. , Porter Megan , Stark Michael J. R. 
Published: May 2005

AbstractThe Kluyveromyces lactis toxin is a heterotrimeric protein which irreversible arrests proliferation of sensitive Saccharomyces cerevisiae cells in the G1 phase of the cell cycle. By expressing the γ subunit of the toxin in sensitive yeast cells from a conditional promoter, it was previously demonstrated that it alone is required for inhibition (Tokunaga et al. (1989). Nucleic Acids Res. 17, 3435–3446). Here we show that, like native exogenous toxin, intracellular γ subunit expression promoters a striking arrest of sensitive cells in G1. However, unlike the G1 arrest caused by native toxin, that induced by the γ subunit alone does not result in reduced cellular viability and is fully and rapidly reversible, suggesting that the G1 arrest and the irreversibility of action may reflect different aspects of the toxin's interaction with sensitive cells. We have selected a large number of S. cerevisiae mutants which are highly resistant to the toxin in order to study its mode of action in more detail. Complementation analysis demonstrated that all but one of the mutants were recessive and these defined four separate genes. Members of two complementation groups concurrently acquired resistance to intracellular γ subunit expression, suggesting that they contain a modified toxin target site. The other two genes appear to be required for entry of the γ subunit into the sensitive cell since these mutants, while refractory to exogenous toxin, were fully sensitive to intracellular γ subunit expression.
Cell wall glucomannoproteins of Saccharomyces cerevisiae mnn9
Authors: Van Rinsum Johanna , Klis Frans M. , Ende Herman Van Den 
Published: May 2005

AbstractMannoproteins were isolated from Saccharomyces cerevisiae mnn9 mutant cell walls by laminarinase digestion and purified by affinity and anion‐exchange chromatography. The purified mannoprotein fraction contained three predominant proteins with molecular masses of 300 kDa, 220 kDa and 160 kDa. These compounds were absent in an SDS extrct of cell walls or in a hot‐citrate extract of mnn9 cells.The carbohydrate part of the purified mannoproteins consisted of (N‐acetyl)glucosamine, mannose and glucose in a molar ratio of 1:53:4. O‐Glycosidically linked chains, containing 70% of the mannose, were released by mild β‐elimination. N‐Glycosidically linked chains, representing 80% of the (N‐acetyl)glucosamine and 20% of the mannose, were released by peptide N‐glycosidase F (PNGase F) digestion. Complete degradation of protein by alkaline hydrolysis released besides the N‐ and O‐glycosidically linked chains, another type of carbohydrate chain containing the residual (N‐acetyl)glucosamine, mannose and most of the glucose in a molar ratio of 1:17:18. Glucose was β‐glycosidically linked.The results indicate that β‐glucose is linked to PNGase F‐resistant N‐linked chains present on cell wall mannoproteins. We propose that these chains are responsible for the linkage between mannoproteins and glucan in the cell wall.
Purification and some properties of membrane‐bound and soluble pyrophosphatases of yeast vacuoles
Authors: Lichko Lidiya , Okorokov Lev 
Published: May 2005

AbstractThe membrane‐bound and soluble pyrophosphatase (PPase) activities of Saccharomyces carlsbergenis vacuoles are determined by the functioning of special enzymes and are not due to non‐specific PPi hydrolysis by other vacuolar phosphohydrolases. The molecular mass of the membrane‐bound PPase is apparently 120 000 and its molecule consists of three subunits with Mr = 41 000. Soluble PPase has a molecular mass of about 82 000 and includes three subunits with Mr = 28 000. Both enzymes are glycoproteins. The vacuolar membrane‐bound PPase is a proton pump.
Structure of the yeast endoplasmic reticulum: Localization of ER proteins using immunofluorescence and immunoelectron microscopy
Authors: Preuss Daphne , Mulholland Jon , Kaiser Chris A. , Orlean Peter , Albright Charles , Rose Mark D. , Robbins Phillips W. , Botstein David 
Published: May 2005

AbstractThe endoplasmic reticulum (ER) and other secretory compartments of Saccharomyces cerevisiae have biochemical functions that closely parallel those described in higher eukaryotic cells, yet the morphology of the yeast organelles is quite distinct. In order to associate ER functions with the corresponding cellular structures, we localized several proteins, each of which is expected to be associated with the ER on the basis of enzymatic activity, biological function, or oligosaccharide content. These marker proteins were visualized by immunofluorescence or immunoelectron microscopy, allowing definition of the S. cerevisiae ER structure, both in intact cells and at the ultrastructural level. Each marker protein was most abundant within the membranes that envelop the nucleus and several were also found in extensions of the ER that frequently juxtapose the plasma membrane. Double‐labeling experiments were entirely consistent with the idea that the marker proteins reside within the same compartment. This analysis has permitted, for the first time, a detailed characterization of the ER morphology as yeast cells proceed through their growth and division cycles.
A high‐affinity uptake system for branched‐chain amino acids in Saccharomyces cerevisiae
Authors: Tullin Søren , Gjermansen Claes , Kielland‐Brandt Morten C. 
Published: May 2005

AbstractIn order to isolate mutants with impaired uptake of branched‐chain amino acids, mutants were induced which on complex medium were sensitive to an inhibitor of branched‐chain amino acid biosynthesis. Eighteen mutants of independent origin were found. Ten of them were assayed for branched‐chain amino acid uptake. Three of these were impaired in the uptake of L‐valine, L‐isoleucine and L‐leucine, while the rest were unaffected in uptake of any of the three amino acids. Kinetics of the uptake by one selected mutant and the parental strain S288C were compared to models for one or two systems obeying Michaelis‐Menten kinetics. This analysis suggested that a high‐affinity system for all three amino acids is absent in the mutant, whereas low‐affinity uptake of L‐isoleucine and L‐leucine by one or more systems remains unaffected. Moreover, medium‐affinity uptake components for L‐valine and L‐leucine, not originally seen in the wild type, were identified in the mutant. In the wild type, 10 mM of any of the amino acids L‐alanine, L‐cysteine, L‐isoleucine, L‐leucine, L‐tryptophan and L‐valine reduce uptake of any of the three branched‐chain amino acids. We propose that a permease responsible for high‐affinity uptake of the branched‐chain amino acids in strain S288C is partially or completely inactive in the mutant. Tetrad analysis shows that the phenotype can be ascribed to a single Mendelian gene. The wild‐type allele is denoted BAP1 for branched‐chain amino acid permease. The BAP1‐dependent system is different from the general amino acid permease.
A rapid method for localized mutagenesis of yeast genes
Authors: Muhlrad Denise , Hunter Rosalyn , Parker Roy 
Published: May 2005

AbstractWe have developed a simple procedure for the localized mutagenesis of yeast genes. In this technique the region of interest is first amplified under mutagenic chain reaction (PCR) conditions. Cotransformation of the PCR product with a gapped plasmid containing homology to both ends the PCR procuct allows in vivo recombination to repair the gap with the mutagenized DNA. This procedure is efficient, allows trageting of specific regions for mutagenesis, and requires no subcloning steps in Escherichia coli.
IMP2, a nuclear gene controlling the mitochondrial dependence of galactose, maltose and raffinose utilization in Saccharomyces cerevisiae
Authors: Donnini Claudia , Lodi Tiziana , Ferrero Iliana , Puglisi Pier Paolo 
Published: May 2005

AbstractThe IMP2 gene of Saccharomyces cerevisiae is involved in the nucleo‐mitochondrial control of maltose, galactose and raffinose utilization as shown by the inability of imp2 mutants of grow on these carbon sources in respiratory‐deficient conditions or in the presence of ethidium bromide and erythromycin. The negative phenotype cannot be scored in the presence of inhibitors of respiration and oxidative phosphorylation, indicating that the role of the mitochondria in the utilization of the above‐mentioned carbon sources in imp2 mutants is not at the energetical level.Mutantions in the IMP2 gene also confer many phenotypic alterations in respiratory‐sufficient conditions, e.g. leaky phenotype on oxidizable carbon sources, sensitivity to heat shock and sporulation deficiency.The IMP2 gene has been cloned, sequenced and disrupted. The phenotype of null imp2 mutants is indistinguishable from that of the originally isolated mutant.
Identification of a Saccharomyces cerevisiae homolog of the SNF2 transcrioptional regulator in the DNA sequence of an 8·6 kb region in the LTE1‐CYS1 interval on the left arm of chormosome I
Authors: Clark Michael W. , Zhong Wu Wei , Keng Teresa , Storms Reginald K. , Barton Arnold , Kaback David B. , Bussey Howard 
Published: May 2005

AbstractThe DNA sequence of an 8·6 kb region of the left arm of chromosome I has been determined. This region, between the LTEL and CYS1 loci, is approximately 40 kb from centromere. There are six potential open‐reading frames (ORFs), Provisionally nemed YAL001‐006 within this fragment of chromosome I. Four of these ORFs can be aligned with Previously indentified FUN transcripts: FUN28 with YAL006, FUN29 with YAL004, FUN30 with YAL001 and FUN31 with YAL002. The YAL001 ORF shows significant homology to the SNF2 transcriptional regulator. A region of the DNA contains an extensive repeat of the bases C‐A‐T positioned in the 5′ terminus of the YAL004 promoter region.
Sequence of the sup61‐RAD18 region on chromosome III of Saccharomyces cerevisiae
Authors: Benit Paule , Chanet Roland , Fabre Francis , Faye Gerard , Fukuhara Hiroshi , Sor Frederic 
Published: May 2005

AbstractA 7965 bp DNA segment from the right arm of chromosome III of Saccharomyces cerevisiae, encompassing the sup61 and RAD18 genes, was sequenced. Four new open reading frames were found in this DNA fragment. One of them YCR103, is 51% homologous with the G10 gene product of Xenopus laevis.
Architectural features of pre‐mRNA introns in the fission yeast Schizosaccharmyces pombe
Authors: Prabhala Ganesh , Rosenberg George H. , Käufer Norbert F. 
Published: May 2005

AbstractThe architectural features of 73 introns found in 36 genes of the fission yeast Schizosaccharomyces pombe have been compiled and tabulated. The intron features of Saccharomyces cerevisia and other eukaryotes. The results that S. pombe displays quite different architectural features than the budding yeast S. cerevisiae. However, particularly in the 3′ region, S. pombe introns also appear to differ from mammalian introns.
The complete sequence of K3B, a 7·9 kb fragment between PGK1 and CRY1 on chromosome III, reveals the presence of seven open reading frames
Authors: Bolle Paul‐André , Gilliquet Véronique , Berben Gilbert , Dumont Jacques , Hilger François 
Published: May 2005

AbstractWe have determined the nucleotide sequence of a segment from chromosome III fo Saccharomyces cerevisiae extending over 7·9 kb between the PGK1 and CRY1 loci.The fragment contains seven open reading frames, YCR241, YCR242, YCR243, YCR244, YCR245, YCR246 and YCR247, of more than 70 codons. The study of the effects of a global disruption of YCR242, YCR243, YCR244, YCR245 and YCR247 shows that they are not essential for growth division.
The peroxisomal import signal of amine oxidase from the yeast Hansenula polymorpha is not universal
Authors: De Hoop M. J. , Valkema R. , Kienhuis C. B. M. , Hoyer M. A. , Ab G. 
Published: May 2005

AbstractAmine oxidase from the yeast Hansenula polymorpha is a peroxisomal protein. The signal for routing of the protein into peroxisomes has not been identified yet. Expression of a mutant amine oxidase in H. Polymorpha has revealed that the C‐terminal sequence, which possesses an internal SRL tripeptide, is not involved in targeting (Faber et al., unpublished). We have explored heterologous expression of the amine oxidase gene (AMO) in Saccharomyces cerevisiae to investigate the conservation of peroxisomal targeting pathways between yeasts. Surprisingly, wide‐type amine oxidase is not recognized as a peroxisomal protein by S. cerevisiae. The enzyme, which was fully active and acumulated to levels similar to those found in H. polymorpha, stayed entirely in the cytosol. However, fusing a SKL or a SRL sequence to the C‐terminus forced the protein at least partially into peroxisomes of the heterologous host. These data suggest that the functional targeting sequence of amine oxidase may differ from the C‐terminal peroxisomal targeting signal S/C/A‐K/R/H‐L (Gould et al., 1989). Contrary to the established tripeptide motif, the amine oxidase targeting signal appears not to be conserved between the different yeast species.
Mutations in phosphofructokinases alter the control characteristics of glycolysis in vivo in Saccharomyces cerevisiae
Authors: Lloyd David , James Christopher J. , Maitra Pabitra K. 
Published: May 2005

AbstractEthanol and CO2 production from gluecose by non‐proliferating suspensions of aerobicaly‐grown, glucose‐derepressed wild‐type Sacharomyces cerevisiae is inhibited by O2; monitoring by mass spectrometry provides a direct method for measurement of the Pasteur effect.Under aerobic conditons, that part of the CO2 evolved equivalent to the O2 consumed, is produced by respiration: subtraction of this respiratory CO2 from the total gives, CO2 produced by aerobic glycolysis. Pasteur quotients (anaerobic CO2/aerobic glycolytic CO2) were within the range 1.2 to 3.0. The Pasteur effect was not observed in the presence of carbonyl cyanid m‐chlorophenylhydrazone, an uncoupler of mitochondrial energy metabolism, or in a ρ cytoplasmic petite mutant. A ‘non‐allosteric’ mutant with an altered regulatory subunit of phosphofructokinase showed no Pasteur effect. Strains bearing a nonsense mutation pfk1 in the catalytic subnit of soluble phosphofructokinase (PFKI) also showed no Pasteur effect; the residual fermentative activity of this strain was dependent on PFKII, the particulate phosphofructokinase. A double mutant lacking both PFKI and glucose‐6‐phosphat dehydrogenase showed similar characteristics to those of the single pfk1 mutant; this indicates that the hexose monophosphate shunt is not acting to bypass the phosphofructokinase block. A ‘hyper‐allosteric’ mutant altered in the regulatory subunit encoded by the gene PFK2 showed characteristics of glucose fermentation and ethanol oxidation very similar to those of wild‐type organisms. These results indicate that either of the two phosphofructokinases can cary out glycolysis.
A Ser/Thr‐rich multicopy suppressor of a cdc24 bud emergence defect
Authors: Bender Alan , Pringle John R. 
Published: May 2005

AbstractMSB2 was identified previously as a multicopy suppressor of a temerature‐sensitive mutation in CDC24, a gene required for polarity establishment and bud formation in Saccharomyces cerevisiae. The inferred MSB2 product contains 1306 amino acids, 42% of which are Ser or Thr. Its Ser+Thr‐richnes and hydrophobicity profile suggest that Msb2p may be an integral membrane protein containing a long, periplasmic, N‐terminal domain and a short, cytoplasmic, C‐terminal domain. Cells that lack MSB2 display no obvious mutant phenotypes. MSB2 is located between the centromere and KSS1 on the right arm of chromosome VII. Although physical mapping suggests that MSB2 and LEU1 (on the left arm of chromosome VII) are approximately 40 kb apart, the genetic map distance observed between leul and msb2 :: URA3 marker was only 2.3 cM.
Isolation and sequence analysis of the gene encoding translation elongation factor 3 from Candida albicans
Authors: Di Domenico B. J. , Lupisella J. , Sandbaken M. , Chakraburtty K. 
Published: May 2005

AbstractThe structural gene encoding translation elongation factor 3 (EF‐3) has been cloned from a Candida albicans genomic library by hybrization to a Saccharomyces cerevisiae probe containing the Saccharomyces gene, YEF3 (Sandbaken et al., 1990b). The sequences were shown to be functionally homologous to the Saccharamyces gene by three criteria: (1) a Saccharomyces strain transformed with a high copy plasmid containing CaEF3 sequences overprodues the EF‐3 peptide two‐fold; (2) extracts from this strain exhibit a two‐fold increase in the Ef‐3 catalysed, ribosome‐dependent ATPase activity (Kamath and Chakraburtty, 1988); and (3) the Candida gene complements a Saccharomyces null mutant. The coding region, identified by DNA sequencing, indicates that CaEF3 encodes a 1050 amino acid polypeptide having a potential molecular weight of 116 865 Da. This protein shows 77% overall identity to the Saccharomyces YEF3 gene, with a significantly greater identity (94%) concentrated in the region of the protein thought to contain the catalytic domain of EF‐3 (Sandaken et al., 1990a). The upstream non‐coding region contains T‐rich regions typical of many yeast genes and several potential RAPI/GRFI elements shown to regulate expression of a number of translational genes (Mager, 1988). The data confirm a high degree of conservation for EF‐3 among the two organisms.
Molecular analysis of yeast chromosome II between CMD1 and LYS2: The excision repair gene RAD16 located in this region belongs to a novel group of double‐finger proteins
Authors: Mannhaupt Gertrud , Stucka Rolf , Ehnle Susanne , Vetter Irene , Feldmann Horst 
Published: May 2005

AbstractWe have analysed a region some 30 kb centromere distal form PHO5 on the right arm of yeast chromosome II and determined the nucleotide sequence of a 8.95 kb DNA segment from this region. By this analysis we were able to derive the precise location and the transcriptional orientation of CMD1, ALG1, SSN6 and LYS2. An open reading frame of 2370 bp was locatlized between SSN6 and LYS2, which has recently been identified (Schild et al., 1991) to be the RAD16 gene. The putative gene product, 790 amino acids in length, reveals several interesting freatures. It contains a nuclear target singnature and shares several blocks of similarity with the yeast recombinational repair protein RAD54 and the nuclear factor SNF2 (SW12), which is required for teh transcriptioal activation of a number of yeast genes. The similarity blocks in these three proteins are reminiscent of those found in the helicase superfamily. Furthrmore, RAD16 contains a novel ‘double‐finger’ motif, which has been encountered in a variety of proteins from different organisms that are suggested to interact with DNA and are involved in diverse functions including site‐specific recombination, DNA repair, and transcriptional regulation. The putative gene product of RAD16 then is the first example of a proteins in which the novel double‐finger motif is found to be combined with a poteintial DNA helicase framework.
Efficient selection of phleomycin‐resistant Saccharomyces cerevisiae transformants
Authors: Wenzel Thibaut. J. , Migliazza Anna , Yde Steensma H. , Van Den Berg Johan A. 
Published: May 2005

AbstractThe recently dsecribed dominant yeast marker Tn5ble confers phleomycin resistance on the yeast Saccharomyces cerevisiae (Gatignol, Baron and Tiraby, 1987. Mol. Gen. Genet. 207, 342–348). Incubation in non‐selective medium prior to selection is critical, however, for getting phleomycin‐resistant transformants. A 6‐h incubation period was found to give optimal transformation frequencies, up to 105 transformants/μg plasmid, comparable to selection for uracil prototrophy (Ura+).
Antibodies directed against a yeast carboxyl‐terminal peroxisomal targeting signal specifically recognize peroxisomal proteins from various yeasts
Authors: Aitchison John D. , Szilard Rachel K. , Nuttley William M. , Rachubinski Richard A. 
Published: May 2005

AbstractThe carboxyl‐terminal tripeptide Ala‐Lys‐Ile is essential for targeting Canadida tropicalis trifunctional enzyme (hydratase‐dehydrogenase‐epimerase) to peroxisomes of both Candida albicans and Saccharomyces cerevisiae (Aitchison, J. D., Murray, W. W. and Rachubinski, R. A. (1991). J. Biol. Chem. 266, 23197–23203). We investigated the possibility that this tripeptide may act as a general peroxisomal targeting signal (PTS) for other proteins in the yeasts C. tropicalis, C. albicans, Yarrowia lipolytica and S. cerevisiae, and in rat liver. Anti‐AKI antibodies raised against the carboxyl‐terminal 12 amino acids of trifunctional enzyme were used to search for this PTS in proteins of these yeasts and of rat liver. The anti‐AKI antibodies reacted exclusively with multiple peroxisomal proteins from the yeasts C. tropicalis, C. albicans and Y. lipolytica. There was a weak reaction of the antibodies with one peroxisomal protein from S. cerevisiae and no reaction with peroxisomal proteins from rat liver. Antibodies directed against a synthetic peptide containing a carboxyl‐terminal Ser‐Lys‐Leu PTS (Gould, S. J., Krisans, S., Keller, G.‐A. and Subramani, S. (1990). J. Cell Biol. 110, 27–34) reacted with multiple peroxisomal proteins of rat liver and with peroxisomal proteins of yeast distinct from those identified with anti‐AKI antibodies. These results provide evidence that several peroxisomal proteins of different yeasts contain a PTS antigenically similar to that of C. tropicalis trifunctional enzyme and that this signal is absent from peroxisomal proteins from at least one mammalian system, rat liver.
III. Yeast sequencing reports. Nucleotide sequence of 9·2 kb left of CRY1 on yeast chromosome III from strain ab972: Evidence for a ty insertion and functional analysis of open reading frame YCR28
Authors: Carbone M. L. Agostoni , Panzeri L. , Falconi M. Muzi , Carcano C. , Plevani P. , Lucchini G. 
Published: May 2005

AbstractWe report the 9210 bp sequence from a segment of yeat chromosome III cloned from strain AB972 in λPM3270. Analysis of this sequence and its comparison with the one derived from the corresponding segment of strain XJ24‐24A revealed that the AB972 region contains a duplication of about 2 kb and a Ty element, which are not found in XJ24‐24A and cause a quite significant rearrangement of the whole region. We performed analysis of YCR28, the largest open reading frame we found in both AB972 and XJ24‐24A. YCR28 encodes a putative protein of 512 amino acids with some similarities to yeast allontoate permease. Its disruption does not cause any detectable phenotype on rich medium or on allantoate medium, while we observed a strain‐dependent effect on senstivity to amino acid balance and to 3‐aminotriazole, when cells were grown in synthetic medium.