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  • 1
    ISSN: 1432-0983
    Keywords: Key wordsSaccharomyces ; Tup1-Ssn6 repression ; IME1 ; Sin4-Rgr1 repression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ime1 plays a pivotal role in the initiation of meiosis in a/α diploid cells of Saccharomyces cerevisiae. In the absence of glucose and nitrogen, IME1 expression is greater in a/α cells than in either a or α cells and therefore only a/α, but not a/a or α/α, cells are committed to sporulation. It is known that IME1 expression is positively regulated by Mck1, Rim1, Ime4 and the Swi-Snf complex but other factors may also be involved. In addition, Rme1 is assumed to repress IME1 expression. To provide more details of the repression of expression of IME1, we have isolated mutants in which the IME1p-PHO5 fusion gene integrated at the ura3 locus is expressed in α cells under nutritionally rich conditions. We found that mutations occurred in TUP1, SSN6, SIN4 and RGR1, among which TUP1 and SSN6 were identified for the first time as negative regulators of IME1 expression. Deletion of the Rme1-binding site from the IME1 promoter did not result in activation of the expression of IME1 under nutritionally rich conditions, suggesting that Rme1 does not function as a DNA-binding protein with the Tup1-Ssn6 repression complex. We also demonstrated that the 294-bp fragment from nucleotide position –914 to –621 and the 301-bp fragment from nucleotide position –1215 to –915 of the IME1 promoter region contain elements acting as URS and UAS in TUP1 + and tup1 mutant cells, respectively. These findings indicate that IME1 is negatively regulated by the Tup1-Ssn6 repressor complex through two distinct upstream regions in conjunction with unidentified DNA-binding proteins.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Key words Homeodomain protein ; Mating-type control ; Matα2 ; Disulfide bond ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Matα2 homeodomain protein plays a pivotal role in the control of cell type in Saccharomyces cerevisiae. The homeodomain in the C-terminal region of Matα2 functions as a DNA-binding domain and the N-terminal region, containing two cysteine residues at positions 33 and 34, is thought to be involved in formation of Matα2 homodimers via disulfide bonds. matα2 mutants, isolated in a previous study, in which haploid-specific genes cannot be repressed by the Mata1-Matα2 heterodimer but a-specific genes can be repressed by the Matα2 homodimer, were found to produce mutant Matα2 with a substitution of tyrosine or phenylalanine for Cys33. To clarify the role of Cys33 and Cys34 in the Matα2 protein, we generated several matα2 mutants by site-directed mutagenesis which had serine residues in place of these Cys residues. Transforming MAT a cells with plasmids carrying these matα2 (MATα1 +) mutations rendered transformants unable to mate. Northern blot analysis revealed that transcription of the a-specific gene STE2 and the haploid-specific locus RME1 in these transformants is repressed to the same level as in wild-type MAT a/MATα cells. We concluded that neither Cys33 nor Cys34 is required for repression of a-specific genes by the Matα2 homodimer or of haploid-specific genes by the Mata1-Matα2 heterodimer, and therefore suggest that Matα2 homodimer formation in vivo is not mediated by disulfide linkage.
    Type of Medium: Electronic Resource
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