bluuebunny/crossref_metadata_2025_split huggingface dataset:
Production of senescent cells of Saccharomyces cerevisiae by centrifugal elutriation
Authors: Woldringh C. L. , Fluiter K. , Huls P. G. 
Published: May 2005

AbstractThe centrifugal elutriator has been used as a baby machine by loading the chamber with a population of mixed‐generation daughter cells and allowing this population to grow, divide and age under continuous washing‐out of newborn daughter cells. Clear peaks in the number of elutriated cells were reproducibly obtained for at least ten generations. The parent cells growing in the chamber continued to divide at the steady‐state generation time of 95–100 min, showing no change in cycle time during aging. The washed‐out daughter cells increased in volume during the first five generations from their steady‐state value of 17 μm3 to a maximum of 34 μm3. As to be expected, the generation times of these large daughters, determined in a synchronous batch culture, were shorter (130 min) than that of the steady‐state daughters (240 min), even when derived from 15‐generation parents. No indication for a volume increase of daughter cells without bud was observed when a population was allowed to grow in the chamber without washing‐out the smaller daughter cells. The 15‐generation parent population, recovered from the chamber, had an average volume of 80 μm3 and consisted of: (i) 71% cells with more than ten scars, (ii) 13% cells with one to nine scars, and (iii) 17% daughter cells. The production of senescent cells by undisturbed growth in the elutriator chamber has been prolonged to 29 generations. The method is therefore suitable to examine what factors determine the life span of budding yeast.
The complete sequence of a 9037 bp DNA fragment of the right arm of Saccharomyces cerevisiae chromosome VII
Authors: Arroyo Javier , García‐Gonzalez Melba , García‐Saez M. Isabel , Sánchez Miguel , Nombela César 
Published: May 2005

AbstractWe report the sequence of a 9037 bp fragment from the right arm of Saccharomyces cerevisiae chromosome VII. Analysis of the sequence revealed four complete open reading frames (ORFs), namely G7572, G7576, G7579 and G7584. The first three corresponded, respectively, to the previously cloned genes: HIP1, coding for a high‐affinity histidine‐specific permease, TDH1, one of the known genes coding for glyceraldehyde‐3‐phosphate dehydrogenase and ODPX, which encodes a precursor of protein X, a component of the pyruvate dehydrogenase complex. The ORF G7584 showed 35·8% identity with a hypothetical protein of Caenorhabditis elegans chromosome 3. The reported sequence has been deposited in the EMBL data library under Accession Number X82408.
Two‐Dimensional protein map of Saccharomyces cerevisiae: Construction of a gene–protein index
Authors: Boucherie Helian , Monribot Christelle , Perrot Michel , Dujardin Genevieve , Kermorgant Michele , Slonimski Piotr 
Published: May 2005

AbstractThis publication marks the beginning of the construction of a gene–protein index that relates proteins which are resolved on the two‐dimensional protein map of Saccharomyces cerevisiae with their corresponding genes. We report the identification of 36 novel polypeptide spots on the yeast protein map. They correspond to the products of 26 genes. Together with the polypeptide spots previously identified, this raises to 41 the number of genes whose products have been identified on the protein map. The proteins identified here are concerned with four major areas of yeast cellular physiology: carbon metabolism, heat shock, amino acid biosynthesis and purine biosynthesis. Given the molecular weight and isoelectric point of the identified proteins, and the codon‐usage bias of the corresponding genes, it can be estimated that 25 to 35% of all the soluble yeast proteins are detectable under the labelling and running gel conditions used in this study.
In vivo cloning by homologous recombination in yeast using a two‐plasmid‐based system
Authors: Degryse Eric , Dumas Bruno , Dietrich Mireille , Laruelle Laurence , Achstetter Tilman 
Published: May 2005

AbstractIn order to reduce the number of classical DNA manipulation and ligation steps in the generation of yeast expression plasmids, a series of vectors is described which facilitate the assembly of such plasmids by the more efficient ‘recombination in vivo’ technique. Two sets of vectors were developed. The first set, called ‘expression vectors’, contains an expression cassette with a yeast promoter and the PGK terminator separated by a polylinker, and an Escherichia coli replicon. Subcloning in these vectors of a DNA fragment generates a ‘transfer vector’ which is compatible with the second set of E. coli–yeast shuttle vectors. This set of ‘recombination vectors’ contains a cassette for a functional copy of a gene complementing a host strain auxotrophy or a bacterial gene conferring an antibiotic resistance to the plasmid‐bearing host. Plasmid copy numbers can be modulated through the use of URA3 or URA3‐d as the selective marker together with an ARS/CEN and the 2 μm replicon.Integration of the cloned DNAs into the yeast linearized replicative vectors occurs by recombination between homologous flanking sequences during transformation in yeast or E. coli. All the vectors contain the origin of replication of phage f1 and allow the generation of single‐stranded DNA in E. coli for sequencing or site‐directed mutagenesis.The sequence presented (Figure 1a) has been entered in the EMBL data library under Accession Number Z48747.
Host cell properties and external pH affect proinsulin production by Saccharomyces yeast
Authors: Kozlov Dmitry G. , Prahl Natalija , Efremov Boris D. , Peters Lars , Benevolensky Sergei V. , Wambut Rolf , Karpychev Igor V. , Eldarov Mikhail A. 
Published: May 2005

AbstractThe expression of a hybrid gene encoding an α‐factor prepro leader peptide‐miniproinsulin (MPI) fusion [MPI is the same as the LysArg human insulin precursor described by Thim et al. (1986)] was tested in a series of isogenic yeast strains to investigate the influence of some genetic and physiological factors on heterologous production in yeast. We found that: (i) an MFα1 gene disruption in haploid cells, as well as MFα1 gene product expression in diploid cells, do not affect the MPI secretion level; (ii) under conditions of exogenous leucine availability, MPI production is hindered by leucine auxotrophy (a leu2 mutation); (iii) rho− mutations increase the per‐cell MPI yield approximately three‐fold; (iv) the MPI yield is apparently dependent on the pH of the culture medium: the higher the external pH, the larger the per‐cell MPI yield.
Sequence, map position and genome organization of the RPL17B gene, encoding ribosomal protein L17b in Saccharomyces cerevisiae
Authors: Berroteran Rhonda W. , Hampsey Michael 
Published: May 2005

AbstractSequence analysis of the newly defined SSU81 gene revealed an adjacent open reading frame (ORF) encoding a protein whose deduced amino acid sequence is identical to that of ribosomal protein L17. The DNA sequence of this region is different from that of the RPL17A gene and therefore represents a duplicate gene encoding L17. We have designated this gene RPL17B. The RPL17B coding region is split by an intron that occurs in the same position (codons 14/15) as the intron in RPL17A. The RPL17B promoter region includes two TATA boxes, a canonical UASRPG motif, and several pyrimidine‐rich tracts. RPL17B was mapped by CHEF and lambda clone grid hybridization blots to the right arm of chromosome V, linked to the TRP2 and RAD51 genes. A partial ORF was identified adjacent to RPL17B and SSU81 that is homologous to an ORF (designated A509) physically linked to RPL17A. This observation, and the identical position of the introns within the RPL17 genes, suggest that one RPL17 locus arose by duplication and translocation of the other. The complete 3·8 kbp DNA sequence encompassing RPL17B has been entered in the GenBank data library under Accession Number U15653.
The Saccharomyces cerevisiae FLO1 flocculation gene encodes for a cell surface protein
Authors: Bidard Frederique , Bony Muriel , Blondin Bruno , Dequin Sylvie , Barre Pierre 
Published: May 2005

AbstractThe sequencing of a 6619 bp region encoding for a flocculation gene previously cloned from a strain defined as FLO5 (Bidard et al., 1994) has revealed that it was a FLO1 gene. The FLO1 gene product has been localized at the cell surface of the yeast cell by immunofluorescent microscopy. The Flo1 protein contains four regions with repeated sequences which account for about 70% of the amino acids of this protein. A functional analysis of the major repeated region has revealed that it plays an important role in determining the flocculation level. A gene disruption experiment has shown that the FLO5 strain STX 347‐1D contains at least two flocculation genes of the FLO1 type but that they are supposed to be inactive and do not contribute to its flocculation. However, enzyme‐linked immunosorbent assays performed on intact cells have revealed that a protein expressed at the cell surface of the FLO5 strain STX 347‐1D is antigenically related to Flo1p. A deletion analysis of the 5′ region of the FLO1 gene has shown that the expression is submitted to controls which depend on the genetic background of the strain.
A new essential gene located on Saccharomyces cerevisiae chromosome IX
Authors: Kurlandzka Anna , Rytka Joanna , Gromadka Robert , Murawski Marcin 
Published: May 2005

AbstractA new 1150 amino acids long open reading frame (ORF), coding for an essential protein of unknown function was found Saccharomyces cerevisiae by sequencing 3754 bp of geonomic DNA. The clone was isolated in a search for a fatty acid‐binding protein (FABP) and was localized on chromosome IX. The ORF bears no homology to FABP, but it shows weak similarity to Plasmodium vivax reticulocyte binding protein 1 and to aggregation‐specific adenylate cyclase from Dictyostelium discoideum. The new gene is constitutively transcribed regardless of the carbon source used. The nucleotide sequence reported in this paper has been deposited in GenBank (Accession Number U17918).
Improvement of chromosomal DNA extraction from different yeast species by analysis of single preparation steps
Authors: Cardinali Gianluigi , Pellegrini Leandro , Martini Alessandro 
Published: May 2005

AbstractA modified procedure is proposed for chromosomal DNA extraction based on a cell‐wall lytic enzyme never applied before in pulsed field gel electrophoresis. Protoplasting efficiency is retained under very challenging conditions for enzyme activity, such as those required for non‐Saccharomyces yeasts often characterized by cell walls highly resistant to lysis.
XV. Yeast sequencing reports. Sequence analysis of a 9873 bp fragment of the left arm of yeast chromosome XV that contains the ARG8 and CDC33 genes, a putative riboflavin synthase beta chain gene, and four new open reading frames
Authors: Casas Celia , Aldea Marti , Herrero Enrique , Casamayor Antonio , Ariño Joaquin , Lafuente Maria Jose , Gamo Francisco‐Javier , Gancedo Carlos 
Published: May 2005

AbstractThe DNA sequence of a 9873 bp fragment located near the left telomere of chromosome XV has been determined. Sequence analysis reveals seven open reading frames. One is the ARG8 gene coding for N‐acetylornithine aminotransferase. Another corresponds to CDC33, which codes for the initiation factor 4E or cap binding protein. The open reading frame AOE169 can be considered as the putative gene for the Saccharomyces cerevisiae riboflavin synthase beta chain, since its translation product shows strong homology with four prokaryotic riboflavin synthase beta chains. The nucleotide sequence reported here has been submitted to the EMBL data library under the Accession Number X84036.
XV. Yeast sequencing reports. Sequence analysis of a 44 kb DNA fragment of yeast chromosome XV including the Ty1‐H3 retrotransposon, the suf1(+) frameshift suppressor gene for tRNA‐Gly, the yeast transfer RNA‐Thr‐1a and a delta element
Authors: Vandenbol Micheline , Durand Patrick , Portetelle Daniel , Hilger Francois 
Published: May 2005

AbstractWe have sequenced on both strands a 44,019 bp fragment located on the left arm of Saccharomyces cerevisiae chromosome XV.The sequenced segment contains 22 open reading frames (ORFs) of at least 100 amino acids long, one of which probably contains an intron. Six of the 22 ORFs correspond to known proteins: the multicopy suppressor of Snf1 protein 1, the two Ty1‐H3 transposon proteins TyA and TyB, the myo‐inositol transporter 2, the transcription factor protein Ino4 and the 3,4‐dihydroxy‐5‐hexaprenylbenzoate methyltransferase. Of the 16 remaining ORFs, two show highest homologies with the yeast serine/threonine protein kinase Ste20 and the human tryptophanyl‐tRNA synthetase. Eight ORFs show slight similarities with protein sequences described in data banks.DNA sequence comparison reveals also the presence of three known sequences: the Ty1‐H3 transposable element, the yeast suf1(+) frameshift suppressor gene for tRNA‐Gly and the yeast transfer RNA‐Thr‐1a. A fourth DNA sequence shows striking identities with the yeast delta elements.The 44,019 bp sequence has been entered in the EMBL data library under Accession Number Z48149.
A disruption‐replacement approach for the targeted integration of foreign genes in Hansenula polymorpha
Authors: Agaphonov Michael O. , Beburov Michael Yu. , Ter‐Avanesyan Michael D. , Smirnov Vladimir N. 
Published: May 2005

AbstractA system has been developed which allows the selection of integrative transformants with replacement of the Hansenula polymorpha methanol oxidase gene (MOX) with expression cassettes carrying heterologous gene under the control of the MOX promoter. The system is convenient for comparison of the expression levels of different constructs integrated into the same locus of the H. polymorpha genome. This system was used to compare the secretion levels of human urinary plasminogen activator, the secretion of which was directed by different signal sequences.
XV. Yeast sequencing reports. DNA sequence analysis of a 13 kbp fragment of the left arm of yeast chromosome XV containing seven new open reading frames
Authors: Casamayor Antonio , Aldea Martí , Casas Celia , Herrero Enrique , Gamo Francisco‐Javier , Lafuente María J. , Gancedo Carlos , Ariño Joaquín 
Published: May 2005

AbstractThe sequence of a 13 kbp fragment located in the vicinity of the left telomere of chromosome XV (cosmid pEOA179) has been determined. Seven new open reading frames (ORFs) encoding polypeptides longer than 100 residues have been found (AOB629, AOA342, AOC231, AOE555, AOE236, AOA236 and AOE1045). Three of them show no identity with proteins deposited in the data banks. ORF AOB629 (629 amino acids) has some similarity with previously described ferric reductases from Saccharomyces cerevisiae and Schizosaccharomyces pombe. ORF AOA342 encodes a polypeptide reminiscent of dihydroflavonol‐4‐reductases from a number of plant species. AOE236 displays a high level of identity when compared with peroxisomal membrane proteins previously cloned from the methylotrophic yeast Candida boidinii. Finally, AOE1045 encodes a large protein (1045 residues) with some identity with a hypothetical 147 kDa protein identified during the sequencing of Caenorhabditis elegans chromosome 3. The complete nucleotide sequence of the 13 kbp fragment has been deposited at the EMBL data base (Accession Number Z48239).
Yeast mapping reports. Clone bank of the mitochondrial genome of yeast Hansenula wingei
Authors: Sekito Takayuki , Okamoto Kozi , Kitano Hiromichi , Yoshida Kazuo 
Published: May 2005

AbstractFor sequencing, mitochondrial DNA from Hansenula wingei yeast was digested with various restriction enzymes and the resultant DNA fragments were cloned into a pEMBL phasmid vector. Our clone bank consists of 39 overlapping clones which cover the entire 27 694 bp region of the H. wingei mitochondrial genome. The nucleotide sequence data reported in this paper will appear in the DDBJ, EMBL and GenBank Nucleotide Sequence Database with the following Accession Number: D31785.
Further definition of the sequence and position requirements of the arginine control element that mediates repression and induction by arginine in Saccharomyces cerevisiae
Authors: Crabeel Marjolaine , De Rijcke Martine , Seneca Sara , Heimberg Harry , Pfeiffer Ilona , Matisova Andrea 
Published: May 2005

AbstractRepression or induction of the genes involved in arginine biosynthesis or catabolism, respectively, both require participation of the ArgRp/Mcm1p regulatory complex. Our previous work showed that those opposite effects were mediated by a similar arginine‐responsive element of 23 nucleotides (that we now call ARC, for ARginine Control) situated close to the start of transcription in the repressed promoters and far upstream of the TATA‐element in the induced promoters.To define more precisely the sequence and position requirements of the ARC element, we have now characterized by mutagenesis the promoter elements of the arginine‐repressible ARG1 and ARG8 genes. We also identify a functional ARC in the CPA1 promoter, thereby confirming, in agreement with our previous mRNA pulse‐labelling data, the participation of a transcriptional component in the arginine regulation of that gene otherwise submitted to a translational regulation.From the 12 ARC elements now characterized, we have derived a consensus sequence and show that such a synthetic element is able to mediate ArgRp/Mcm1p‐dependent arginine regulation.An important new finding illustrated by ARG1 and CPA1, is that contrary to what all the previous data suggested, repression can be mediated by ARC elements located far upstream of the TATA‐box. The new data suggest that the arginine repressor might inhibit transcription in an active process.
Physiological and molecular characterization of flor yeasts: Polymorphism of flor yeast populations
Authors: Martínez P. , Codón A. C. , Pérez L. , Benítez T. 
Published: May 2005

AbstractYeast strains which form velum on the surface of Sherry wine during the aging process have been isolated and characterized. According to their metabolic and molecular features most of the yeasts that were isolated belong to different races of Saccharomyces cerevisiae (beticus, cheresiensis, montuliensis and rouxii). Due to the conditions under which these yeasts were isolated, all strains have in common the capacity to develop a film as an adaptive mechanism which allows them to grow and survive in 15·5% vol. ethanol. All strains were prototrophs for amino acids and most vitamins but they gave different responses to the killer factor. However, whereas their physiological features were similar, they showed a great heterogeneity with regards to the nuclear and mitochondrial genome (mtDNA): DNA content per cell was quite variable (1·3 to 2n), electrophoretic karyotypes of nuclear genomes indicated a main pattern with some variations, and polymorphism shown by the mtDNA was very high. Under extreme conditions such as Sherry wine with 15·5% vol. ethanol, no fermentable sugar and an exclusively oxidative metabolism, cells hardly grow and the maintenance of a live population depends on survival and respiration, which in turn depend on the mtDNA. At the same time these environmental conditions are mutagenic for the mtDNA, causing an increase in variation. Thus, the polymorphism observed might reflect the enormous variability induced by the ethanol followed by the selection of those mtDNA sequences which make the mitochondria metabolically active under these conditions.
Carvedilol improves left ventricular function in murine coxsackievirus‐induced acute myocarditis Association with reduced myocardial interleukin‐1β and MMP‐8 expression and a modulated immune response
Authors: Pauschinger Matthias , Rutschow Susanne , Chandrasekharan Kumaran , Westermann Dirk , Weitz Anneke , Schwimmbeck Lothar Peter , Zeichhardt Heinz , Poller Wolfgang , Noutsias Michel , Li Jun , Schultheiss Heinz‐Peter , Tschope Carsten 
Published: May 2005

BackgroundProinflammatory cytokines induce the expression of matrix metalloproteinases that play a crucial role in myocardial remodeling. Beta‐adrenergic receptor stimulation influences the production of cytokines heralding the possibility of modulating cytokine production by beta‐adrenergic blockers.Methods and resultsIn a coxsackievirus B3 murine myocarditis model (BALB/c), effects of carvedilol and metoprolol on myocardial cytokine expression, inflammatory cell infiltration and MMP/TIMP profiles were investigated. In carvedilol‐treated mice, a significant improvement in left ventricular function was documented 10 days post infection. In infected mice (n=10), IL‐1β, TNF‐α, TGF‐β1 and IL‐10 myocardial mRNA abundance were increased significantly (240%, 200%, 161%, and 230%) compared to controls (n=10), while IL‐15 mRNA was markedly reduced (70%). Infected mice showed significantly increased infiltrations with CD3‐, CD4‐ and CD8‐T‐lymphocytes (730%, 1110%, 380%). In the infected mice, myocardial MMP/TIMP profiles presented a significant upregulation of membrane type‐1 MMP, MMP‐9, MMP‐8 and MMP‐3 (150%, 160%, 340%, and 270%) and a significant decrease in TIMP‐4 levels (75%). Carvedilol attenuated over‐expression of myocardial TGF‐β1, IL‐1β and MMP‐8 mRNA expression significantly and induced a relevant IL‐10 mRNA expression in the infected mice (n=10). By an unchanged infiltration with CD3‐T‐lymphocytes, carvedilol showed a representative reduction in CD4‐T‐lymphocytes.ConclusionCarvedilol treatment in experimental myocarditis leads to reduced expression of proinflammatory cytokines and MMPs, which contributes to reduced matrix degradation and ultimately to improved structural integrity of the heart. Besides the antiadrenergic potential, carvedilol is beneficial due to a wide range of biological activities (antiinflammatory, antifibrotic, antioxidative and immunomodulatory).
The uptake of D-araboascorbic acid (D-isoascorbic acid) by guinea-pig tissues
Authors: Hughes R. E. , Hurley R. J. 
Published: May 2005

1. Guinea-pigs on a low, controlled intake of ascorbic acid (0.3 mg/100 g body-weight) were given daily an oral supplement of L-ascorbic acid (AA) or of D-isoascorbic acid (isoAA) (both 1.5 mg/100 g body-weight). The AA was deposited in the tissues, the isoAA was not.2. Intramuscularly administered isoAA was deposited in the tissues but not to the same extent as equivalent amounts of intramuscularly administered AA.3. Paper chromatography confirmed that intramuscularly administered isoAA was deposited in the tissues as isoAA, i.e. there was no conversion into AA.4. It is concluded that in guinea-pigs (a)AA is absorbed from the gastro-intestinal tract much more readily than isoAA and (b)AA is either more readily abstracted from the blood or is more readily retained by the tissues or both, than is isoAA.