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  • 1
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    Oxford : Oxford University Press
    Call number: 04-Zell:409a
    Keywords: Saccharomyces cerevisiae / Cytology ; Schizosaccharomyces pombe / Cytology ; Cell nuclei ; Fungal molecular biology
    Pages: xix, 316 p. : ill. (some col.)
    ISBN: 0199637733
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  • 2
    ISSN: 1432-072X
    Keywords: Anthranilate synthase ; Cell permeabilisation ; Indoleglycerolphosphate synthase ; Saccharomyces cerevisiae ; Tryptophan biosynthetic enzymes ; Tryptophan pool
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The free tryptophan pool and the levels of two enzymes of tryptophan biosynthesis (anthranilate synthase and indoleglycerolphosphate synthase) have been determined in a wild type strain of Saccharomyces cerevisiae and in mutants with altered regulatory properties. The tryptophan pool of wild type cells growing in minimal medium is 0.07 μmole per g dry weight. Addition of anthranilate, indole or tryptophan to the medium produces a fifteen- to forty-fold increase in tryptophan pool, but causes no repression of the biosynthetic enzymes. Inclusion of 5-methyltryptophan in the growth medium causes a reduction in growth rate and a derepression of the biosynthetic enzymes, and this is shown here not to be correlated with a decrease in the free tryptophan pool. Mutants with an altered anthranilate synthase showing decreased sensitivity to inhibition by l-tryptophan or by the analogue dl-5-methyltryptophan have a tryptophan pool far higher than the wild type strain, but no repression of indoleglycerolphosphate synthase was observed. Mutants with an anthranilate synthase more sensitive to tryptophan inhibition show a slightly reduced tryptophan pool, but no derepression of indoleglycerolphosphate synthase was found. A mutant with constitutively derepressed levels of the biosynthetic enzymes shows a considerably increased tryptophan pool. Addition of 5-methyltryptophan to the growth medium of non-derepressible mutants causes a decrease in growth rate accompanied by a decrease in the tryptophan pool.
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  • 3
    ISSN: 1432-0983
    Keywords: Benomyl resistance ; Yeast ; Mitosis ; Cell cycle mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated 150 benomyl resistant mutants of the fission yeast Schizosaccharomyces pombe. Seven of these mutants were found to be cold sensitive for mitosis. These mutants were the subject of physiological, cytological and genetical characterisation. Growth and division of the seven mutants were similar to the wild type strain at 35 °C. After shift from the permissive (35 °C) to the restrictive temperature (20 °C) the mutants became blocked in mitosis whilst cellular growth continued. Consequently, elongate cells were formed. Six of the seven benomyl resistant mutants became blocked in mitosis at 20 °C with a single aberrant nucleus. In every case the benomyl resistant and cold sensitive phenotype was due to a mutation in a single nuclear gene. These mutants were found to comprise a single genetic linkage group (ben4) and were unlinked to existing TBZ/MBC resistant mutants of S. pombe. Whilst no cross resistance was found in our mutants to TBZ, six of the seven mutants were super sensitive to the spindle poison CIPC. We believe that the phenotype exhibited by these mutants is consistent with a defective tubulin subunit.
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  • 4
    ISSN: 1432-0983
    Keywords: Mitotic control ; Schizosaccharomyces pombe ; Cell size ; wee mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The major cell cycle control in the fission yeast Schizosaccharomyces pombe acts at entry to mitosis, and involves three previously identified genes cdc2, cdc25 and wee1. The presence of a weel mutation phenotypically suppresses cdc25 mutations. This paper describes the isolation and subsequent analysis of a strain in which the suppression is reversed by the presence of a new mutation, designated win1.1. The mutation causes a slight increase in cell size at division in most genetic backgrounds. However, when combined with a wee1 mutation and cdc25.22, the win 1.1 mutation interacts strongly to generate a novel phenotype: cells are phenotypically cdc during growth on minimal medium but cdc + when cultured on complex medium. The win1 locus is unlinked to previously identified genes involved in mitosis.
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  • 5
    ISSN: 1432-0983
    Keywords: Key words Cell cycle ; DNA replication ; DNA polymerase ; Fission yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Highly purified DNA polymerase δ from the fission yeast Schizosaccharomyces pombe is a complex of at least four distinct subunits. Genes encoding three of these (pol3 + /cdc6 + , cdc1 + and cdc27 + ) have been characterised previously. Here we describe the isolation and characterisation of cdm1 + , the gene encoding the smallest (22kDa) subunit of the Pol δ complex. Over-expression of cdm1 + , which encodes a 160 amino-acid protein with no significant sequence similarity to proteins in current databases, is able to rescue cells carrying temperature-sensitive mutations in either pol3 + /cdc6 + , cdc1 + or cdc27 + . Cells deleted for cdm1 + are viable, indicating that cdm1 + is non-essential for mitotic growth, and are no more sensitive to a variety of DNA replication inhibitors and DNA damaging agents than are wild-type cells. In addition, over-expression of cdm1 + suppresses the temperature-sensitive cdc24-M38 mutant suggesting that cdc24 + may also have a role in DNA polymerase δ function.
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  • 6
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Fig. 1 Cross-hybridization of SPT16 to DNA from a variety of species. Total DNA from the species indicated was digested with an excess of the restriction endonuclease EcoRl (3 units per /ug, 37 °C, overnight). The amount of DNA loaded on to a 0.8% agarose gel was ~10g, except for the D. ...
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  • 7
    ISSN: 1617-4623
    Keywords: cdc2 ; Cell cycle ; Mitotic control ; Schizosaccharomyces pombe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Five new elements of the mitotic control in the fission yeast Schizosaccharomyces pombe were isolated from gene libraries as multicopy suppressors of the conditional lethal phenotype of win1-1 weel ts cdc25ts triple mutant strains. These genes were designated wisl + −wis5+for win suppressing, and do not correspond to winl + or any of the previously characterised mitotic control genes. None of the wis genes is capable of suppressing the cdc phenotype of cdc25 ts strains, suggesting that their effect is not simply to reverse the effect of loss of cdc25 function. wisl + has been previously reported to encode a putative serine/threonine protein kinase that acts as a dosage-dependent inducer of mitosis. wis4 + appears to be a specific suppressor of the winl-1 mutation. wis2 + and wis3 + are capable of suppressing a wide range of cdc phenotypes arising from the combination of various mutations with wee1 ts and cdc25 ts, suggesting that the wis2 + and wis3 + products may interact with elements central to the mitotic control.
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  • 8
    ISSN: 1617-4623
    Keywords: Cell cycle ; Mitosis ; Fission yeast ; cdc27 +
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cell division cycle gene cdc27 + of the fission yeast Schizosaccharomyces pombe is required for the transition from G2 into mitosis. Genetic and physiological experiments suggest a close relationship between cdc27 + and the cdc2 + gene, a key regulator of mitosis in yeast and also in higher eukaryotic cells. We isolated the cdc27 + gene by complementation of a temperature-sensitive cdc27 mutant. The DNA sequence of this gene predicts a 1116 nucleotide open reading frame split by five short introns, ranging in size from 49 to 74 nucleotides. Analysis of cDNA clones confirmed the structure of the gene. The deduced cdc27 + gene product consists of 372 amino acids with a predicted Mr of 43 kDa. No homology of the predicted protein with known proteins could be found, thus the cdc27 + gene encodes a novel function required for the G2-M transition. Northern analysis revealed two mRNAs of 1.4 and 2.2 kb transcribed from this gene, the smaller transcript being approximately tenfold more abundant than the larger. The level of cdc27 + mRNAs remained constant through the cell cycle indicating that the time of action of the cdc27 + gene, which is known to be regulated by elements of the mitotic control, is not determined by periodic accumulation of its transcripts.
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  • 9
    ISSN: 1617-4623
    Keywords: Cell cycle ; Fission yeast ; Schizosaccharomyces pombe ; Ribonucleotide reductase ; DNA replication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cdc22 + gene of Schizosaccharomyces pombe is required early in the cell cycle, and its transcript varies in concentration in step with the cell cycle, with a peak level at the G1-S boundary. The sequences of the cdc22 + gene and of a multicopy suppressor of cdc22 ts mutations, suc22 +, have been determined. The cdc22 + open reading frame, which is interrupted in the genome by a single intron very close to its 5′ end, encodes a protein of 811 amino acids, which has an amino acid sequence highly similar to that of the large subunit of ribonucleotide reductase from several species. The suc22 + gene contains an uninterrupted open reading frame of 391 amino acids, very similar to the sequence of the small subunit of ribonucleotide reductase. Disruption of either gene is lethal. Upstream of the cdc22 + coding region are seven short sequence elements similar to the recognition sequence for MluI, which are involved in regulating periodic transcription of the gene. Inhibition of DNA synthesis by hydroxyurea results in a several-fold increase in the level of the cdc22 + transcript. In contrast, hydroxyurea does not induce the 1.5 kb transcript of suc22 +, but results in the induction of a 1.9 kb mRNA which hybridises to suc22 + DNA.
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  • 10
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] We initially established that Mlul motifs (Figs la and 20) are physiologically significant in fission yeast. A plasmid containing three synthetic Mlul sequences linked to the Escherichia coli lacZ gene was transformed into a wild-type strain and the cell-cycle expression of the lacZ ...
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