Localization and properties of a silencing element near the mat3-M mating-type cassette of Schizosaccharomyces pombe.
Transcription is repressed in a segment of Schizosaccharomyces pombe chromosome II that encompasses the mat2-P and mat3-M mating-type cassettes. Chromosomal deletion analysis revealed the presence of a repressor element within 500 bp of mat3-M. This element acted in synergy with the trans-acting factors Swi6, Clr1, Clr2, Clr3, and Clr4 and had several properties characteristic of silencers: it did not display promoter specificity, being able to silence not only the M mating-type genes but also the S. pombe ura4 and ade6 genes placed on the centromere-distal side of the mat3-M cassette; it could repress a gene when placed further than 2.6 kb from the promoter and it acted in both orientations, although with different efficiencies, the natural orientation repressing more stringently than the reverse. Following deletion of this element, two semistable states of expression of the mat3-M region were observed and these two states could interconvert. The deletion did not affect gene expression in the vicinity of the mat2-P cassette, 11 kb away from mat3-M. Conversely, deleting 1.5 kb on the centromere-proximal side of the mat2-P cassette, which was previously shown to partially derepress transcription around mat2-P, had no effect on gene expression near mat3-M. A double deletion removing the mat2-P and mat3-M repressor elements had the same effect as the single deletions on their respective cassettes when assayed in cells of the M mating type. These observations allow us to refine a model proposing that redundant pathways silence the mating type region of S. pombe.
Thon
G
G
Department of Genetics, Institute of Molecular Biology, University of Copenhagen, DK-1353 Copenhagen K, Denmark. gen@biobase.dk
Bjerling
K P
KP
Nielsen
I S
IS
D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D018797:Cell Cycle Proteins / Q000235:genetics; D002503:Centromere / Q000235:genetics; D003433:Crosses, Genetic; D004926:Escherichia coli / Q000235:genetics; D005800:Genes, Fungal / Q000502:physiology*; D049770:Genes, Mating Type, Fungal; D005838:Genotype; D008954:Models, Biological; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010641:Phenotype; D010957:Plasmids; D012568:Schizosaccharomyces / Q000235:genetics*; D014170:Transformation, Genetic
D018797:Cell Cycle Proteins