A Synthetic silencer mediates sir-dependent functions in Saccharomyces cerevisiae

Francis J. McNally, Jasper Rine

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Abstract

Copies of the mating-type genes are present at three loci on chromosome III of the yeast Saccharomyces cerevisiae. The genes at the MAT locus are transcribed, whereas the identical genes at the silent loci, HML and HMR, are not transcribed. Several genes, including the four SIR genes, and two sites, HMR-E and HMR-I, are required for repression of transcription at the HMR locus. Three elements have been implicated in the function of the HMR-E silencer: a binding site for the RAP1 protein, a binding site for the ABF1 protein, and an 11-bp consensus sequence common to nearly all autonomously replicating sequence (ARS) elements (putative origins of DNA replication). RAP1 and ABF1 binding sites of different sequence than those found at HMR-E were joined with an 11-bp ARS consensus sequence to form a synthetic silencer. The synthetic silencer was able to repress transcription of the HMRa1 gene, confirming that binding sites for RAP1 and ABF1 and the 11-bp ARS consensus sequence were the functional components of the silencer in vivo. Mutations in the ABF1 binding site or in the ARS consensus sequence of the synthetic silencer caused nearly complete derepression of transcription at HMR. The ARS consensus sequence mutation also eliminated the ARS activity of the synthetic silencer. These data suggested that replication initiation at the HMR-E silencer was required for establishment of the repressed state at the HMR locus.

Original languageEnglish (US)
Pages (from-to)5648-5659
Number of pages12
JournalMolecular and Cellular Biology
Volume11
Issue number11
StatePublished - Nov 1991

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Consensus Sequence
Saccharomyces cerevisiae
Binding Sites
Genes
Mutation
Replication Origin
DNA Replication
Protein Binding
Chromosomes
Yeasts
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

A Synthetic silencer mediates sir-dependent functions in Saccharomyces cerevisiae. / McNally, Francis J.; Rine, Jasper.

In: Molecular and Cellular Biology, Vol. 11, No. 11, 11.1991, p. 5648-5659.

Research output: Contribution to journalArticle

McNally, Francis J. ; Rine, Jasper. / A Synthetic silencer mediates sir-dependent functions in Saccharomyces cerevisiae. In: Molecular and Cellular Biology. 1991 ; Vol. 11, No. 11. pp. 5648-5659.
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