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Volumn 8, Issue 2, 1998, Pages 233-239

Mechanisms of silencing in Saccharomyces cerevisiae

Author keywords

[No Author keywords available]

Indexed keywords

AGING; ARTICLE; HETEROCHROMATIN; MATING TYPE; NONHUMAN; NUCLEOLUS; PRIORITY JOURNAL; SACCHAROMYCES CEREVISIAE; SILENCER GENE; YEAST;

EID: 0032055730     PISSN: 0959437X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0959-437X(98)80146-9     Document Type: Article
Times cited : (115)

References (60)
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    • of special interest. Second site-suppressors of cis-acting mutations in the HMRE silencer were identified. One of the suppressors, SAS2, reduced silencing at HML, while enhancing silencing at HMR in an Orc2p- and Orc5p-dependent fashion. Sequencing of the gene revealed significant homology to known acetyltransferases.
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    • Ehrenhofer-Murray, A.E.1    Rivier, D.H.2    Rine, J.3
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    • RLF2, a subunit of yeast chromatin assembly factor-I, is required for telomeric chromatin function in vivo
    • of outstanding interest. Previous studies from this group identified a series of mutations defective in Rap1p localization to discrete peripheral loci. In this study, one of these genes, RLF2, was demonstrated to be identical to CAC1, encoding a sub-unit of chromatin assembly factor-I. Mutations in RLF2 reduced the efficiency of telomeric silencing, while not influencing telomere size. Although Rap1p clustering was eliminated, telomeres remained clustered, suggesting that telomeres can cluster in the absence of high concentrations of Rap1p. The authors propose that CAF-1 rapidly deposits a specific subclass of acetylated histones to form a repressed chromatin state.
    • Enomoto S, McCune-Zierath PD, Gerami-Nejad M, Sanders MA, Berman J. RLF2, a subunit of yeast chromatin assembly factor-I, is required for telomeric chromatin function in vivo. of outstanding interest Genes Dev. 11:1997;358-370 Previous studies from this group identified a series of mutations defective in Rap1p localization to discrete peripheral loci. In this study, one of these genes, RLF2, was demonstrated to be identical to CAC1, encoding a sub-unit of chromatin assembly factor-I. Mutations in RLF2 reduced the efficiency of telomeric silencing, while not influencing telomere size. Although Rap1p clustering was eliminated, telomeres remained clustered, suggesting that telomeres can cluster in the absence of high concentrations of Rap1p. The authors propose that CAF-1 rapidly deposits a specific subclass of acetylated histones to form a repressed chromatin state.
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    • of outstanding interest. A replication-dependent chromatin assembly factor was identified in yeast by biochemical fractionation and shown to consist of three subunits, denoted Cac1p, Cac2p, and Cac3p. Peptides from each of the subunits were sequenced, and the wild-type genes were isolated. Each subunit displayed weak, but significant, homology to its human counterpart. Mutations in any of the subunits lead to UV hypersensitivity and a partial loss of telomeric silencing.
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    • Separable functions of ORC5 in replication initiation and silencing
    • of outstanding interest. Spontaneous revertants of the orc5-1 allele resulted in the identification of novel orc5 alleles that were specifically defective in either replication or in HMR silencing. Mutations that abrogated silencing were identified that were not defective in replication at the HMRE origin of replication, as determined by two-dimensional gel electrophoresis. The presence of both classes of alleles in an orc5-1 mutant strain led to trans-complementation of both defects, further indicating that the role of ORC in silencing and replication may be functionally distinct.
    • Dillon A, Rine J. Separable functions of ORC5 in replication initiation and silencing. of outstanding interest Genetics. 147:1997;1053-1062 Spontaneous revertants of the orc5-1 allele resulted in the identification of novel orc5 alleles that were specifically defective in either replication or in HMR silencing. Mutations that abrogated silencing were identified that were not defective in replication at the HMRE origin of replication, as determined by two-dimensional gel electrophoresis. The presence of both classes of alleles in an orc5-1 mutant strain led to trans-complementation of both defects, further indicating that the role of ORC in silencing and replication may be functionally distinct.
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    • The yeast Cac1 protein is required for the stable inheritance of transcriptionally repressed chromatin at telomeres
    • of special interest. The authors use a single-cell assay to demonstrate that cac1 mutant cells that were initially repressed were incapable of maintaining growth. These data suggest a defect in the inheritability of the repressed state. In contrast, no defect was found in the establishment of telomeric silencing.
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    • Silencing factors participate in DNA repair and recombination in Saccharomyces cerevisiae
    • of special interest. Two-hybrid analysis identified Sir4p as a protein capable of interacting with the 70kDa subunit of the yeast Ku heterodimer, Hdf1p. Like hdf1 mutants, mutations in SIR2, SIR3, and SIR4 were defective in end-to-end joining and in combination with the rad52 mutation were defective in double-strand DNA repair. Epistasis analysis revealed that the Sir proteins and Hdf1p were acting in the same pathway. The authors propose that Hdf1p interacts with Sir4p at double-strand breaks to promote a heterochromatic state amenable to DNA repair.
    • Tsukamoto Y, Kato J-i, Ikeda H. Silencing factors participate in DNA repair and recombination in Saccharomyces cerevisiae. of special interest Nature. 388:1997;900-903 Two-hybrid analysis identified Sir4p as a protein capable of interacting with the 70kDa subunit of the yeast Ku heterodimer, Hdf1p. Like hdf1 mutants, mutations in SIR2, SIR3, and SIR4 were defective in end-to-end joining and in combination with the rad52 mutation were defective in double-strand DNA repair. Epistasis analysis revealed that the Sir proteins and Hdf1p were acting in the same pathway. The authors propose that Hdf1p interacts with Sir4p at double-strand breaks to promote a heterochromatic state amenable to DNA repair.
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    • An unusual form of transcriptional silencing in yeast ribosomal DNA
    • of special interest. Genes cloned into a Ty1 element, when transposed into the rDNA array, are transcriptionally repressed. This effect was independent of the gene tested but completely dependent on its presence in the rDNA array. A single repeat of rDNA containing the transposon placed elsewhere in the genome was not subject to silencing. This process was dependent on Sir2p but not on other Sir proteins. Psoralen is a reagent that photoreacts with accessible DNA upon UV treatment. Psoralen crosslinking analysis of silenced regions was consistent with a more condensed chromatin state dependent on Sir2p.
    • Smith JS, Boeke JD. An unusual form of transcriptional silencing in yeast ribosomal DNA. of special interest Genes Dev. 11:1997;241-254 Genes cloned into a Ty1 element, when transposed into the rDNA array, are transcriptionally repressed. This effect was independent of the gene tested but completely dependent on its presence in the rDNA array. A single repeat of rDNA containing the transposon placed elsewhere in the genome was not subject to silencing. This process was dependent on Sir2p but not on other Sir proteins. Psoralen is a reagent that photoreacts with accessible DNA upon UV treatment. Psoralen crosslinking analysis of silenced regions was consistent with a more condensed chromatin state dependent on Sir2p.
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    • Transcriptional silencing of Ty1 elements in the RDN1 locus of yeast
    • of special interest. Transposition of a HIS3-marked Ty1 element into the rDNA array led to SIR2-dependent silencing of the HIS3 gene. Silencing was dependent on Rad6p/Ubc2p, a ubiquitin-conjugating activity. The domains of Rad6p involved suggested, however, that this process was not mediated by the N-end rule protein degradation pathway. Depletion of the histones H2A and H2B also led to a loss of silencing, linking alterations in chromatin structure with the rDNA-silencing process.
    • Bryk M, Banerjee M, Murphy M, Knudsen KE, Garfinkel DJ, Curcio MJ. Transcriptional silencing of Ty1 elements in the RDN1 locus of yeast. of special interest Genes Dev. 11:1997;255-269 Transposition of a HIS3-marked Ty1 element into the rDNA array led to SIR2-dependent silencing of the HIS3 gene. Silencing was dependent on Rad6p/Ubc2p, a ubiquitin-conjugating activity. The domains of Rad6p involved suggested, however, that this process was not mediated by the N-end rule protein degradation pathway. Depletion of the histones H2A and H2B also led to a loss of silencing, linking alterations in chromatin structure with the rDNA-silencing process.
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    • Redistribution of silencing proteins from telomeres to the nucleolus is associated with extension of life span in S. cerevisiae
    • of outstanding interest. The SIR4-42 allele suppresses uth4 truncation alleles, resulting in extended mother cell lifespan. Interestingly, these cells also display a relocalization of both Sir3p and Sir4-42p to the nucleolus. Using a magnetic-sorting technique that separates young from old mother cells on the basis of the degree of biotin incorporation in the cell surface (previously developed in this lab), the authors demonstrated that Sir3p relocalizes from the telomere to the nucleolus during the aging process.
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    • (1997) Cell , vol.89 , pp. 381-391
    • Kennedy, B.K.1    Gotta, M.2    Sinclair, D.A.3    Mills, K.4    McNabb, D.S.5    Murthy, M.6    Pak, S.M.7    Laroche, T.8    Gasser, S.M.9    Guarente, L.10
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    • Changes of telomere length cause reciprocal changes in the lifespan of mother cells in Saccharomyces cerevisiae
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    • Accelerated aging and nucleolar fragmentation in yeast sgs1 mutants
    • of special interest. The yeast SGS1 gene is homologous to the human gene mutated in Werner's syndrome, one of the symptoms of which is premature aging. To test the role of Sgs1p in yeast aging, the number of generations a mother cell could produce a viable daughter was assayed. Mutants in Sgs1p, which localize to the nucleolus, exhibited a severely truncated lifespan, with a concomitant loss of HM silencing, relocalization of Sir3p to the nucleolus, and enlargement and fragmentation of the nucleoli.
    • Sinclair DA, Mills K, Guarente L. Accelerated aging and nucleolar fragmentation in yeast sgs1 mutants. of special interest Science. 277:1997;1313-1316 The yeast SGS1 gene is homologous to the human gene mutated in Werner's syndrome, one of the symptoms of which is premature aging. To test the role of Sgs1p in yeast aging, the number of generations a mother cell could produce a viable daughter was assayed. Mutants in Sgs1p, which localize to the nucleolus, exhibited a severely truncated lifespan, with a concomitant loss of HM silencing, relocalization of Sir3p to the nucleolus, and enlargement and fragmentation of the nucleoli.
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    • Sinclair, D.A.1    Mills, K.2    Guarente, L.3
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    • Mammalian homologues of the Polycomb-group gene Enhancer of zeste mediate gene silencing in Drosophila heterochromatin and at S. cerevisiae telomeres
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