bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of polygalacturonase, pectin‐lyase and pectin‐esterase activities in a Saccharomyces cerevisiae strain
Authors: Gainvors A. , Frézier V. , Lemaresquier H. , Lequart C. , Aigle M. , Belarbi A. 
Published: May 2005

AbstractThe catalytic capacity of several excreted pectinolytic enzymes obtained from various yeast strains was examined using in vivo and biochemical techniques. Of the 33 yeast strains studied, 30 were isolated from champagne wine during alcoholic fermentation. Only one yeast strain was found to excrete pectinolytic enzymes and was identified as Saccharomyces cerevisiae and designated SCPP. Pulsed‐field gel electrophoresis and the polymerase chain reaction technique were used to characterize further this specific strain. Three types of pectinolytic enzymes were found to be excreted by SCPP: polygalacturonase, pectin‐lyase and pectin‐esterase. These enzymes allow pectin hydrolysis during cell growth.
XIV. Yeast sequencing reports. Twelve open reading frames revealed in the 23·6 kb segment flanking the centromere on the Saccharomyces cerevisiae chromosome XIV right arm
Authors: Verhasselt Peter , Aert Rita , Voet Marleen , Volckaert Guido 
Published: May 2005

AbstractThe nucleotide sequence of 23·6 kb of the right arm of chromosome XIV is described, starting from the centromeric region. Both strands were sequenced with an average redundancy of 4·87 per base pair. The overall G+C content is 38·8% (42·5% for putative coding regions versus 29·4% for non‐coding regions). Twelve open reading frames (ORFs) greater than 100 amino acids were detected. Codon frequencies of the twelve ORFs agree with codon usage in Saccharomyces cerevisiae and all show the characteristics of low level expressed genes. Five ORFs (N2019, N2029, N2031, N2048 and N2050) are encoded by previously sequenced genes (the mitochondrial citrate synthase gene, FUN34, RPC34, PRP2 and URK1, respectively). ORF N2052 shows the characteristics of a transmembrane protein. Other elements in this region are a tRNAPro gene, a tRNAAsn gene, a τ34 and a truncated δ34 element. Nucleotide sequence comparison results in relocation of the SIS1 gene to the left arm of the chromosome as confirmed by colinearity analysis. The nucleotide sequence data reported in this paper will appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the accession number X77395.
Respiratory inhibitors affect incorporation of glucose into Saccharomyces cerevisiae cells, but not the activity of glucose transport
Authors: Walsh Michael C. , Smits Hans‐Peter , van Dam Karel 
Published: May 2005

AbstractIncubation of starved galactose‐grown S. cerevisiae cells with cyanide reduced glucose uptake as measured over a 5‐s period. The Vmax for glucose uptake was decreased by over a factor of two but the apparent affinity for glucose doubled. When measured in the sub‐second time scale, however, there was no significant inhibition of glucose uptake, by cyanide, up to 200‐ms, clearly demonstrating that, in cyanide treated cells, glucose uptake was not linear for the first 5‐s.After a 200‐ms exposure of untreated cells to radio‐labelled glucose, less than 10% of the intracellular label resided in soluble uncharged compounds. In cyanide‐treated cells up to 43% of the labelled compounds were uncharged, with a concurrent reduction of intracellular label residing in anionic compounds. The results suggest that, in the presence of 10 mM cyanide when respiration is inhibited, a reduction in the cellular ATP concentration causes a reduction in hexose‐kinase activity which results in an accumulation of internal free glucose, which in turn causes a reduction in net glucose transport.
Characterization of an active GST‐human Cdc2 fusion protein kinase expressed in the fission yeast Schizosaccharomyces pombe: A new approach to the study of cell cycle control proteins
Authors: Leroy Dorothée , Baldin Véronique , Ducommun Bernard 
Published: May 2005

AbstractCharacterization of cdk (cyclin dependent kinases) substrates and studies of their regulation require purified enzymatic complexes of cdc2‐related catalytic and cyclin regulatory subunits. We produced human Cdc2 kinase in the fission yeast Schizosaccharomyces pombe as a fusion protein with glutathione S‐transferase (GST). The GST‐human Cdc2p fusion protein was active in vivo since it rescued a temperature‐sensitive allele of cdc2. The fusion protein was purified using a one‐step chromatography procedure with glutathione–Sepharose and exhibited a catalytic activity in vitro. Yeast cyclin B and suc1 were found in association with GST‐Cdc2. A 17‐fold stimulation of GST‐Cdc2 kinase activity was obtained by incubation of recombinant human cyclin A with the S. pombe cellular extract prior to affinity purification. This indicates that cyclin concentration is limiting in this overexpression system. These findings describe a fast and easy production of active recombinant human Cdc2 kinase in yeast that can be used for biochemical studies.
The immunodetected yeast purine—cytosine permease is not N‐linked glycosylated, nor are glycosylation sequences required to have a functional permease
Authors: Rodriguez C. , Bloch J. C. , Chevallier M. R. 
Published: May 2005

AbstractThe purine‐cytosine permease (PCP) of the yeast Saccharomyces cerevisiae was detected by immunological methods. Using antibodies directed against synthetic peptides, whose sequences were derived from the primary structure of the PCP, immunoprecipitation of [35S]methionine‐labelled PCP was achieved either from cellular extracts or from in vitro translation mixtures. Non‐labelled PCP was also detected on Western blots of membrane proteins. Similar migration rates were observed for PCP originating both from immunoprecipitated cellular extracts and from in vitro translation mixtures. Hence, post‐translational processing, if any, only slightly affects the size of the protein. Also no evidence was found for N‐linked core‐glycosylation: identical migration rates were observed when immunoprecipitated PCP molecules were extracted from cells labelled for 10 min with [35S]methionine, pretreated or not with tunicamycin.On the other hand, the suppresion of the two potential N‐linked glycosylation sequences in the DNA did not lead to inactivation of the transport activity, confirming that N‐linked glycosylation is not required for the permease activity.
An efficient method to isolate yeast genes causing overexpression‐mediated growth arrest
Authors: Espinet Carme , De La Torre Maria Angeles , Aldea Marti , Herrero Enrique 
Published: May 2005

AbstractIn order to characterize new yeast genes regulating cell proliferation, a number of overexpression‐sensitive clones have been isolated from a Saccharomyces cerevisiae cDNA library in a multicopy vector under the control of the GAL1 promoter, on the basis of growth arrest phenotype under galactose‐induction conditions. Thirteen of the independent clones isolated in this way correspond to previously known genes (predominantly coding for morphogenesis‐related proteins or for multifunctional transcriptional factors), while the remaining 11 independent clones represent new genes with unknown functions. The more stringent conditions employed in this screening compared with previous ones that also employed a dominant genetics approach to isolate overexpression‐sensitive genes has allowed us to extend the number of yeast genes that exhibit this phenotype. The effect of overexpression of MCM1 (whose product participates in the regulation of a number of apparently unrelated cellular functions) has been studied in more detail. Galactose‐induced overexpression of MCM1 leads to rapid growth arrest at the G1 or S cell cycle stages, with many morphologically‐abnormal cells. Several of the other clones also exhibit a G1 arrest terminal phenotype when overexpressed.
In vivo evidence for non‐universal usage of the codon CUG in Candida maltosa
Authors: Sugiyama Hiroki , Ohkuma Moriya , Masuda Yutaka , Park Sun‐Mee , Ohta Akinori , Takagi Masamichi 
Published: May 2005

AbstractAn alkane‐assimilating yeast Candida maltosa had been studied in order to establish systems suitable for biotransformation of hydrophobic compounds. However, functional expression of heterologous genes tested for this purpose had not been successful in several cases. On the other hand, it had been reported that the codon CUG, a universal leucine codon, is read as serine in C. cylindracea. The same altered codon usage had also been suggested by in vitro experiments in some Candida yeasts which are phylogenetically closely related to C. maltosa.In this study we have shown that the failure in functional expression of a heterologous gene is due to the fact that the codon CUG is read as serine in C. maltosa. This conclusion was drawn from the following experimental results: (1) when a cytochrome P450 gene of C. maltosa containing a CTG codon was expressed in C. maltosa, the corresponding amino acid was found to be serine, and not leucine; (2) a tRNA gene with an almost identical structure to that of the tRNA SerCAG gene of C. albicans could be isolated from the genome of C. maltosa; (3) the Saccharomyces cerevisiae URA3 gene, which has one CTG codon, could not complement the ura3 mutation of C. maltosa as itself, but when the CTG codon was changed to another leucine codon, CTC, the mutated gene could complement the ura3 mutation.The last result is the first example of succeeding in functional expression of a heterologous gene in Candida species having an altered codon usage by changing the CTG codon in the gene to another codon.The nucleotide sequence datum reported in this paper will appear in the GSDB, DDBJ, EMBL and NCBI nucleotide sequence databases with the Accession Number D26074.
The sequence of a 13·5 kb DNA segment from the left arm of yeast chromosome XIV reveals MER1; RAP1; a new putative member of the DNA replication complex and a new putative serine/threonine phosphatase gene
Authors: Coster Francoise , Van Dyck Luc , Jonniaux Jean‐Luc , Purnelle Benedicte , Goffeau Andre 
Published: May 2005

AbstractThe nucleotide sequence of two adjacent ClaI fragments from the left arm of Saccharomyces cerevisiae chromosome XIV has been determined. Analysis of the 13,520 bp DNA segment reveals nine open reading frames (ORFs) longer than 300 bp. N1302 contains the consensus sequence for a phosphate‐binding loop common to ATP‐ and GTP‐binding proteins and a strictly conserved ‘SRC’ sequence of unknown function present in all accessory proteins of replicative polymerases. N1306 shares homologies with serine/threonine phosphatases. N1310 encodes RAP1 (TUF or SBF‐E), a transcription regulator. N1330 is the MER1 gene required for chromosome pairing and genetic recombination. Two ORFs show no homology with proteins in the databases and no particular features. N1311 is not likely to be expressed as it is located on the complementary strand of N1310. The sequence has been submitted to the EMBL data library under Accession Number X78898.
Construction of a complete genomic library of Saccharomyces cerevisiae and physical mapping of chromosome XI at 3·7 kb resolution
Authors: Thierry Agnès , Gaillon Laurent , Galibert Francis , Dujon Bernard 
Published: May 2005

AbstractA consortium of European laboratories has been organized to systematically sequence the genome of Saccharomyces cerevisiae. As part of the BIOTECH program aimed at sequencing chromosomes XI and II, we have constructed a total genomic library of yeast strain FY1679 (a direct S288C derivative) into cosmid vectors pWE15 and pOU61cos. Primary clones from four independent libraries totalling 190 genome equivalents have been stored at −80°C.A subset of 1939 independent clones (six genome equivalents) was hybridized using purified chromosomes XI and X as probes. A total of 147 chromosome XI‐specific cosmid clones was used to construct the physical map of that chromosome. Mapping methods included a combination of classical bottom‐up strategies (fingerprinting, hybridizations) and a novel top‐down strategy using I‐SceI chromosome fragmentation. The 147 cosmid clones form a unique contig covering the entire chromosome XI (666 kb) with the sole exceptions of the (C1−3A)n repeats of the telomeres. Colinearity of cosmid inserts with yeast DNA was directly verified. A complete EcoRI map of chromosome XI was deduced from partial overlaps of cosmids and used for the sequencing program. Comparison of this map with the genetic map shows unexpected divergences that have been solved by subsequent genetic analysis, yet underline the necessity of independent physical mapping in genome projects.
Recovery of gene function by gene duplication in Saccharomyces cerevisiae
Authors: Bach M. L. , Roelants F. , De Montigny J. , Huang M. , Potier S. , Souciet J. L. 
Published: May 2005

AbstractA prototroph revertant (Rev9) selected from an ATCase− mutant of the URA2 gene containing three nonsense mutations was shown to contain two ATCase coding sequences. We cloned both ATCase coding areas to show that the duplicated locus (dl9) was the only functional one. Its size corresponded roughly to the second half of the URA2 wild‐type gene. Sequence analysis of the 5′ end of dl9 indicated that this duplicated sequence was inserted within the intergenic region close to the MRS3 gene and was transcribed from an unknown promoter divergently from the MRS3 gene. The event leading to the revertant strain Rev9 included a rearrangement that increased the size of chromosome X by about 60 kb. In agreement with such a rearrangement, recombination was undetectable in the vicinity of the locus dl9. Genetic mapping confirms that the MRS3 gene is 2 cM distal to the URA2 gene on the right arm of chromosome X.
Effects of phleomycin‐induced DNA damage on the fission yeast Schizosaccharomyces pombe cell cycle
Authors: Belenguer Pascale , Oustrin Marie‐Louise , Tiraby Gérard , Ducommun Bernard 
Published: May 2005

AbstractThe effect of phleomycin, a bleomycin‐like antibiotic, has been investigated in the fission yeast, Schizosaccharomyces pombe. We report that in response to phleomycin‐induced DNA damage, growth was inhibited and S. pombe cells arrested in the G2‐phase of the cell cycle. DNA repair mutants rad9 and rad17 did not arrest and were hypersensitive to phleomycin. Cell cycle mutants that entered mitosis without monitoring the completion of DNA replication also displayed an increased sensitivity to this DNA‐damaging agent. Thus, phleomycin could be used as a tool in the fission yeast S. pombe model system for the study of DNA damage and cell cycle checkpoints, or as a new selective agent.
UASNTR functioning in combination with other UAS elements underlies exceptional patterns of nitrogen regulation in Saccharomyces cerevisiae
Authors: Rai Rajendra , Daugherty Jon R. , Cooper Terrance G. 
Published: May 2005

AbstractUASNTR, the UAS responsible for nitrogen catabolite repression‐sensitive transcriptional activation of many nitrogen catabolic genes in Saccharomyces cerevisiae, has been previously thought to operate only as a pair of closely related dodecanucleotide sites each containing the sequence GATAA at its core. Here we show that a single UASNTR site is also able to combine with another unrelated cis‐acting element to mediate transcription as well. In one instance the unrelated cis‐acting element was TTTGTTTAC situated upstream of GLN1, while in another the cis‐acting element was the one previously shown to bind the PUT3 protein. When a UASNTR site functions in combination with an unrelated site, the regulatory responses observed are a hybrid consisting of characteristics derived from both the UASNTR site and the unrelated site as well. These observations resolve several significant inconsistencies that have plagued studies focused on elucidation of the mechanisms involved in the global regulation of nitrogen catabolism.
Tandem integration of multiple ILV5 copies and elevated transcription in polyploid yeast
Authors: Mithieux Suzanne M. , Weiss Anthony S. 
Published: May 2005

AbstractAn industrial yeast strain was modified by introducing DNA into brewing yeast such that the derived cells contain only yeast DNA. Thus selectable markers and bacterial sequences are not present in the final strain, making this procedure attractive for the development of generally acceptable brewing yeast. Linear DNA containing the cloned ILV5 gene was introduced into lager yeast along with an unlinked circular bifunctional plasmid containing a dominant resistance marker. Resistant colonies were screened for site‐directed integration of the ILV5 DNA. Candidates were examined by several methods including Southern transfer and polymerase chain reaction. In this way, a strain WM56 was identified containing three tandem copies of ILV5. The amplified ILV5 region is stable during repeated subculturing in the absence of selective pressure. Correspondingly elevated levels of ILV5 transcript in strain WM56 compared to the control (i.e. non‐tandem) parental strain led to increased amounts of encoded acetohydroxyacid reductoisomerase as evidenced by significantly lower diacetyl production. WM56 appears to be identical to the parental strain judged by CHEF, total restriction digestion patterns, and probing, but differs in the ILV5 region of the chromosome. The method is generally applicable to other yeast strains, and if desired, is amenable to iterated cycles of integration to increase the number of copies.
Glucose metabolism, enzymic analysis and product formation in chemostat culture of Hanseniaspora uvarum
Authors: Venturin C. , Boze H. , Moulin G. , Galzy P. 
Published: May 2005

AbstractThe physiology of Hanseniaspora uvarum K5 was studied in glucose‐limited chemostat cultures and upon glucose pulse. Up to a dilution rate of 0·28 h−1, glucose was completely metabolized in biomass and CO2. Above this value, increase in the dilution rate was accompanied by sequential production of metabolites (glycerol, acetate and ethanol) and decrease in cell yield. Similar results were observed upon glucose pulse. From the enzyme activities (pyruvate dehydrogenase, pyruvate decarboxylase, NAD and NADP‐dependent acetaldehyde dehydrogenases, acetyl coenzyme A synthetase and alcohol dehydrogenase) and substrate affinities, the following conclusions were drawn with respect to product formation of cells: (1) pyruvate was preferentially metabolized via pyruvate dehydrogenase, when biomass and CO2 were the only products formed; (2) acetaldehyde formed by pyruvate decarboxylase was preferentially oxidized in acetate by NADP‐dependent aldehyde dehydrogenase; acetate accumulation results from insufficient activity of acetyl‐CoA synthetase required for the complete oxidation of acetate; (3) acetaldehyde was oxidized in ethanol by alcohol dehydrogenase, in addition to acetate production.
Isolation of the gene HEM4 encoding uroporphyrinogen III synthase in Saccharomyces cerevisiae
Authors: Amillet Jean‐Michel , Labbe‐Bois Rosine 
Published: May 2005

AbstractWe have isolated a genomic DNA fragment that complements the yeast temperature‐sensitive cyt mutation, causing respiratory deficiency and accumulation of porphyrins (Sugimura et al., 1966). Partial DNA sequencing of the complementing region and search for similarity in the DNA and protein databases revealed that (1) the gene had been previously isolated by complementation of the mutation ts2326 (Langgut et al., 1986; accession number X04694), and (2) it encodes a protein with 18–23% identity to uroporphyrinogen III synthases from different sources. This enzyme catalyses the fourth step in the heme biosynthetic pathway and we named its gene HEM4. A hem4Δ disruption mutation was constructed which had phenotypes identical to the cyt mutation. Biochemical analysis confirmed the absence of uroporphyrinogen III synthase activity in both hem4Δ and cyt mutant strains.
Sequence analysis of a 5·6 kb fragment of chromosome II from Saccharomyces cerevisiae reveals two new open reading frames next to CDC28
Authors: Baur Sabine , Becker Jûrgen , Li Ziyu , Niegemann Eckhard , Wehner Eugen , Wolter Ralf , Brendel Martin 
Published: May 2005

AbstractThe sequence of a 5653 bp DNA fragment of the right arm of chromosome II of Saccharomyces cerevisiae contains two unknown open reading frames (YBR1212 and YBR1213) next to gene CDC28. Gene disruption reveals both putative genes as non‐essential. ORF YBR1212 encodes a predicted protein with 71% similarity and 65% identity (total polypeptide of 376 aa) with the 378 aa Sur1 protein of S. cerevisiae, while the putative product of ORF YBR1213, which is strongly expressed, has 28% identity with a Lactococcus lactis‐secreted 45 kDa protein and 24% identity with the Saccharomyces cerevisiae AGA1 gene product. The total sequence of the fragment has been submitted to the EMBL databank (accession number X80224).
Cloning and sequencing of the LEU2 homologue gene of Schwanniomyces occidentalis
Authors: Iserentant D. , Verachtert H. 
Published: May 2005

AbstractA gene that complements the leu2 mutation of Saccharomyces cerevisiae has been cloned from Schwanniomyces occidentalis. The gene codes for a protein of 379 amino acids. As expected for a Schwanniomyces gene, it has a high AT content, which is also reflected in the codon usage. The sequence homology with other known leu2 complementing genes is low. The nucleotide sequence of the Schw. occidentalis LEU2 gene has been assigned the Accession Number X79823 SOLEU2 by EMBL.
Sequencing analysis of a 15·4 kb fragment of yeast chromosome XIV identifies the RPD3, PAS8 and KRE1 loci, and five new open reading frames
Authors: Maftahi M. , Nicaud J.‐M. , Levesque H. , Gaillardin C. 
Published: May 2005

AbstractThe DNA sequence of a 15·4 kb region covering the left arm of chromosome XIV from Saccharomyces cerevisiae was determined. This region contains eight open reading frames (ORFs) which code for proteins of more than 100 amino acids. Three ORFs correspond to the RPD3, PAS8 and KRE1 loci, described previously. Three ORFs show limited homology with known proteins: NO330 with the recessive suppressor of secretory defect SAC1, NO325 with YCR094W identified during chromosome III sequencing; whereas NO315 presents a motif conserved in the dnaJ family. Two ORFs (NO320 and NO325) show no homology to known proteins within the databases screened, but NO320 corresponds to a serine‐threonine‐rich protein. The sequence has been entered in the EMBL data library under Accession Number Z46259.
SSU71, encoding the largest subunit of TFIIF, is located on the right arm of chromosome VII in Saccharomyces cerevisiae
Authors: Sun Zu‐Wen , Hampsey Michael 
Published: May 2005

AbstractSSU71 (TFG1) is an essential nuclear gene encoding the largest subunit of the yeast general transcription factor TFIIF. The SSU71 gene was physically mapped to the right arm of chromosome VII, physically linked to QCR9, by hybridization of the cloned gene to CHEF and lambda clone grid blots. This assignment was confirmed by genetic mapping. A search of the nucleotide sequence databases revealed that SSU71 is immediately adjacent to the TYS1 gene, which encodes tRNATyr synthetase. TYS1 was reported previously to lie on chromosome XV based on sequence overlap with the adjacent UBR1 gene. The mapping data reported here established that TYS1 and UBR1 do not lie on chromosome XV; rather the SSU71‐TYS1‐UBR1 gene cluster lies on the right arm of chromosome VII, physically linked to QCR9 and genetically linked to ade3 and ser2.
XVI. Yeast sequencing reports. A new essential gene GCR4 located in the upstream region of GCR1
Authors: Uemura Hiroshi , Jigami Yoshifumi 
Published: May 2005

AbstractA new essential gene of Saccharomyces cerevisiae was found upstream of GCR1. Its cloning and sequencing predict a 280 amino acid protein (32 577 Da). The predicted protein is fairly hydrophobic, and a search of the database did not identify any homologous proteins. A LEU2 disruption at codon 104 was lethal, but disruption at codon 221 showed a temperature‐sensitive conditional growth phenotype. Abnormalities were observed in some glycolytic enzyme levels. The sequence has been submitted to GenBank‐EMBL‐DDBJ under Accession Number D29645.