Histone H3 and H4 N-termini interact with SIR3 and SIR4 proteins: A molecular model for the formation of heterochromatin in yeast
CellPublished 1 February 1995Open access
Andreas Hecht, Thierry Laroche, Sabine Strahl‐Bolsinger, Susan M. Gasser, Michael Grunstein
Citations774
SJR quartileQ1
SJR score22.61
SNIP7.62
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TL;DR
It is shown that the SIR3 and SIR4 proteins interact with specific silencing domains of the H3 and H4 N-termini in vitro, which proposes a model for heterochromatin-mediated transcriptional silencing in yeast, which may serve as a paradigm for other eukaryotic organisms as well.
Abstract
The silent mating loci and chromosomal regions adjacent to telomeres of S. cerevisiae have features similar to heterochromatin of more complex eukaryotes. Transcriptional repression at these sites depends on the silent information regulators SIR3 and SIR4 as well as histones H3 and H4. We show here that the SIR3 and SIR4 proteins interact with specific silencing domains of the H3 and H4 N-termini in vitro. Certain mutations in these factors, which affect their silencing functions in vivo, also disrupt their interactions in vitro. Immunofluorescence studies with antibodies against RAP1 and SIR3 demonstrate that the H3 and H4 N-termini are required for the association of SIR3 with telomeric chromatin and the perinuclear positioning of yeast telomeres. Based on these interactions, we propose a model for heterochromatin-mediated transcriptional silencing in yeast, which may serve as a paradigm for other eukaryotic organisms as well.
Keywords
Biochemistry, Genetics and Molecular Biology
GeneticsA system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
8,794 Citations1989R Sikorski, P. Hieter
A series of yeast shuttle vectors and host strains has been created to allow more efficient manipulation of DNA in Saccharomyces cerevisiae to perform most standard DNA manipulations in the same plasmid that is introduced into yeast.
GeneSingle-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase
5,960 Citations1988Donald B. Smith, Kevin S. Johnson
Plasmid expression vectors have been constructed that direct the synthesis of foreign polypeptides in Escherichia coli as fusions with the C terminus of Sj26, a 26-kDa glutathione S-transferase (GST; EC 2.5.1.18) encoded by the parasitic helminth Schistosoma japonicum.
Nucleic Acids ResearchImproved method for high efficiency transformation of intact yeast cells
3,266 Citations1992Daniel Gietz, Andrew St. Jean +2 more
Further improvements and streamlining of this method are described, yielding reproducibly 10 6 transformants per microgram of vector DNA, a ten fold improvement, and reducing the time required by 1.5 hours.
GeneMultifunctional yeast high-copy-number shuttle vectors
1,683 Citations1992Thomas Christianson, Robert Sikorski +3 more
A set of four yeast shuttle vectors that incorporate sequences from the Saccharomyces cerevisiae 2 mu endogenous plasmid has been constructed, providing high-copy-number counterparts to the current pRS vectors.
Proceedings of the National Academy of SciencesThe two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest.
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A method that detects proteins capable of interacting with a known protein and that results in the immediate availability of the cloned genes for these interacting proteins is described and could be readily extended to mammalian proteins.
CellPosition effect at S. cerevisiae telomeres: Reversible repression of Pol II transcription
1,348 Citations1990Daniel E. Gottschling, Oscar M. Aparicio +2 more
Yeast telomeres exert a position effect on the transcription of nearby genes, an effect that is under epigenetic control as demonstrated by phenotype and mRNA analyses.
CellModifiers of position effect are shared between telomeric and silent mating-type loci in S. cerevisiae
705 Citations1991Oscar M. Aparicio, Barbara Billington +1 more
The distinctions between telomeres and the HM loci suggest a hierarchy of chromosomal silencing in S. cerevisiae, and insights into telomeric structure and function are provided that are likely to apply to many eukaryotes.
CellPurification and cloning of a DNA binding protein from yeast that binds to both silencer and activator elements
637 Citations1987David Shore, Kim Nasmyth
The purification of RAP1 by DNA affinity chromatography and the isolation of its gene from a lambda gt11 genomic library using antibodies raised against the protein suggest that R AP1 may be a transcriptional regulator that can play a role in either repression or activation of transcription, depending upon the context of its binding site.
CellHistone H4 isoforms acetylated at specific lysine residues define individual chromosomes and chromatin domains in Drosophila polytene nuclei
550 Citations1992Bryan M. Turner, Andrew J. Birley +1 more
Findings support the hypothesis that H4 molecules acetylated at particular sites mediate unique and specific effects on chromatin function.
Genes & DevelopmentEvidence that a complex of SIR proteins interacts with the silencer and telomere-binding protein RAP1.
548 Citations1994Paolo Moretti, Kalev Freeman +2 more
It is proposed that RAP1 acts in the initiation of transcriptional silencing by recruiting a complex of SIR proteins to the chromosome via protein-protein interactions and is consistent with a model in which SIR3 and SIR4 play a structural role in the maintenance of silent chromatin.
Molecular and Cellular BiologyTwo DNA-binding factors recognize specific sequences at silencers, upstream activating sequences, autonomously replicating sequences, and telomeres in Saccharomyces cerevisiae.
509 Citations1988Andrew R. Buchman, William J. Kimmerly +2 more
Genes & DevelopmentSilent domains are assembled continuously from the telomere and are defined by promoter distance and strength, and by SIR3 dosage.
457 Citations1993Hubert Renauld, Oscar M. Aparicio +4 more
Results suggest that a gene's promoter is a key determinant in controlling silencing on that gene and that SIR3 is a crucial component of the silent chromatin domain that initiates at the telomere and is assembled inwardly along the yeast chromosome.
Proceedings of the National Academy of SciencesEthidium bromide provides a simple tool for identifying genuine DNA-independent protein associations.
453 Citations1992J S Lai, Winship Herr
CellExtremely conserved histone H4 N terminus is dispensable for growth but essential for repressing the silent mating loci in yeast
438 Citations1988Paul S. Kayne, U. J. Kim +4 more
Surprisingly, removal of the H4 N terminus also derepresses the silent mating type loci, HML alpha and HMRa, disrupting mating, and this activation is specific since other regulated genes are repressed and induced normally in these cells.
CellSIR3 and SIR4 proteins are required for the positioning and integrity of yeast telomeres
392 Citations1993Francesca Palladino, Thierry Laroche +4 more
It is shown that the SIR3 and SIR4 gene products have a sub-nuclear localization similar to the telomere-associated RAP1 protein, which is found primarily in foci at the nuclear periphery of fixed yeast spheroplasts.
ScienceGenetic Analysis of Histone H4: Essential Role of Lysines Subject to Reversible Acetylation
347 Citations1990Paul C. Megee, Brian A. Morgan +2 more
The structure-function relations of the amino-terminal domain of yeast histone H4 were examined by the creation of directed point mutations and provide genetic proof for the roles of the H4 amino- terminal domain lysines in gene expression, replication, and nuclear division.
Proceedings of the National Academy of SciencesGenetic evidence for an interaction between SIR3 and histone H4 in the repression of the silent mating loci in Saccharomyces cerevisiae.
343 Citations1990Lianna M. Johnson, Paul S. Kayne +2 more
It is shown here that four positions of histone H4 (N-terminal residues 16, 17, 18, and 19) are crucial to silencing and provides a link between one of the SIR proteins and a component of chromatin.
CellYeast histone H4 N-terminal sequence is required for promoter activation in vivo
332 Citations1991Linda K. Durrin, Randall K. Mann +2 more
It is postulate that the H4 N-terminus may interact with a component of the transcription initiation complex, allowing nucleosome unfolding and subsequent initiation.
Proceedings of the National Academy of SciencesThe activation domain of transcription factor PU.1 binds the retinoblastoma (RB) protein and the transcription factor TFIID in vitro: RB shows sequence similarity to TFIID and TFIIB.
312 Citations1993Christian Hagemeier, Andrew J. Bannister +2 more
Evidence that the cellular transcription factor PU.1 can bind to both RB and TFIID is presented and sequence similarity between these two proteins is found, finding that the previously defined domain A of the RB pocket shows sequence similarity to the conserved C terminus of TFIIDs, whereas domain B shows sequence similarities to a second general transcription factor, TFIIB.
The Journal of Cell BiologyLocalization of RAP1 and topoisomerase II in nuclei and meiotic chromosomes of yeast
292 Citations1992Franz Klein, Thierry Laroche +4 more
In interphase nuclei, again in contrast to anti-topoII, anti-RAP1 gives a distinctly punctate staining that is located primarily at the nuclear periphery, suggesting that telomeres are grouped together, perhaps through interaction with the nuclear envelope.
Microbiological ReviewsSilencers, silencing, and heritable transcriptional states
290 Citations1992P Laurenson, Jasper Rine
Analysis of the transcriptional repression at the silent mating-type loci of Saccharomyces cerevisiae may lend insight into heritable repression in other eukaryotes.
Genes & DevelopmentRAP1 and telomere structure regulate telomere position effects in Saccharomyces cerevisiae.
282 Citations1993G Kyrion, Kui Liu +2 more
The data presented here indicate that loss of the carboxyl terminus of RAP1 abolishes position effects at yeast telomeres and diminishes silencing at the HML locus, and it is shown that telomere position effects are highly sensitive to changes in the size (or structure) of the telomeric tract.
Genes & DevelopmentSaccharomyces telomeres assume a non-nucleosomal chromatin structure.
272 Citations1992James H. Wright, Daniel E. Gottschling +1 more
The termini of chromosomal DNA molecules in yeast are assembled in a non-nucleosomal structure encompassing the entire terminal C1-3A tract, separated from adjacent nucleosomes by a region of DNA that is highly accessible to enzymes.
NatureRole of DNA replication in the repression of silent mating type loci in yeast
250 Citations1984Allan M. Miller, Kim Nasmyth
It is shown here that DNA replication is required for the onset of repression, which is necessary for the repression of silent mating type loci in yeast.
NatureHistone H3 amino terminus is required for telomeric and silent mating locus repression in yeast
227 Citations1994Jeffrey S. Thompson, Xuefeng B. Ling +1 more
It is reported that histone H3 is required for full repression at yeast telomeres and at partially disabled silent mating loci, and that the acetylatable lysine residues of H3 play an important role in silencing.
The EMBO JournalEffects of histone H4 depletion on the cell cycle and transcription of Saccharomyces cerevisiae.
227 Citations1988U. J. Kim, Mei Han +2 more
A yeast strain is constructed in which the sole histone H4 gene is under control of the GAL1 promoter and it was found that the resultant lethality manifests itself first in S phase, the period of nucleosome assembly, but leads to highly synchronous arrest in G2 and a virtually complete block in chromosomal segregation.
CellTargeting of SIR1 protein establishes transcriptional silencing at HM loci and telomeres in yeast
205 Citations1993Cheng‐Ting Chien, Stephen W. Buck +2 more
The EMBO JournalHistone H3 N‐terminal mutations allow hyperactivation of the yeast GAL1 gene in vivo.
186 Citations1992Randall K. Mann, Michael Grunstein
The H3 and H4 N‐termini have different functions in both the repression of the silent mating loci and in the regulation of GAL1, as well as a limited extent glucose repression is also alleviated by H3 N‐ terminal deletions.
Genes & DevelopmentRAP1 protein activates and silences transcription of mating-type genes in yeast.
170 Citations1991Stephen E. Kurtz, David Shore
Analysis of rap1ts strains provides evidence that RAP1 functions in both transcriptional activation and silencing of mating-type genes, and several observations indicate that rap1TS strains are defective in the expression of MAT alpha, whose upstream activation sequence contains a R AP1-binding site.
Molecular and Cellular BiologyNotes: Point Mutations in the Yeast Histone H4 Gene Prevent Silencing of the Silent Mating Type Locus <i>HML</i>
166 Citations1990Eun-Chung Park, Jack W. Szostak
The results support an active role for histone H4 in the silencing of gene expression at this locus and a mutation that changed the fourth lysine to alanine resulted in specific derepression of the silent mating type locus HML.
BioEssaysPosition effect variegation in <i>Drosophila</i>: Towards a genetics of chromatin assembly
134 Citations1989Joel C. Eissenberg
The purpose in this review is to summarize some recent work on the genetics of heterochromatin assembly in Drosophila and a recent model for how chromosomal proteins may interact to form aheterochromatic structure.
NatureA highly basic histone H4 domain bound to the sharply bent region of nucleosomal DNA
126 Citations1988Konstantin K. Ebralidse, S. A. Grachev +1 more
It is reported that the amino-acid residue of histone H4 cross-linked at the 1.5 site is histidine-18, embedded in a highly basic cluster Lys-Arg-His- Arg-Lys-Val-Leu-Arg which is probably involved in the sharp bending of the DNA double helix at the ±1 sites.
The EMBO JournalIdentification of a non‐basic domain in the histone H4 N‐terminus required for repression of the yeast silent mating loci.
120 Citations1992Lianna M. Johnson, G. Fisher-Adams +1 more
It is proposed that the basic and non‐basic domains together form a DNA (or protein) induced amphipathic alpha‐helix required in the formation of a repressive chromatin structure.
Molecular and Cellular BiologyPoint mutations in the yeast histone H4 gene prevent silencing of the silent mating type locus HML.
120 Citations1990E C Park, J W Szostak
The results support an active role for histone H4 in the silencing of gene expression at this locus and a mutation that changed the fourth lysine to alanine resulted in specific derepression of the silent mating type locus HML.
Molecular and Cellular BiologyFunctional domains of SIR4, a gene required for position effect regulation in Saccharomyces cerevisiae.
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CellYeast histone H2A and H2B amino termini have interchangeable functions
87 Citations1986Tillman Schuster
This work has shown that switching portions of the N termini between H2B and H2A to create two chimeric histone proteins within the same cell has no obvious effect on viability, which supports the argument that the N-terminal end of one protein complements the function of the other.
GeneticsHISTONE GENE MULTIPLICITY AND POSITION EFFECT VARIEGATION IN <i>DROSOPHILA MELANOGASTER</i>
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The Journal of Cell BiologyA single histone acetyltransferase from Tetrahymena macronuclei catalyzes deposition-related acetylation of free histones and transcription-related acetylation of nucleosomal histones.
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It is argued strongly that two distinct, biologically important histone acetylations, one deposition related and one transcription related, are carried out by a single acetyltransferase.
Cold Spring Harbor Symposia on Quantitative BiologyHistones and the Regulation of Heterochromatin in Yeast
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DevelopmentMechanisms for the construction and developmental control of heterochromatin formation and imprinted chromosome domains
47 Citations1990Kenneth D. Tartof, Marilyn Bremer
A simple mechanism of X-chromosome inactivation is proposed, a purpose for genomic imprinting is suggested, and a general means for regulating the time in development at which certain genes become heterochromatically repressed is postulated.
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A key step in understanding DNA replication has now been reached, in part, through the study of silencing, a process that establishes and maintains repressed transcriptional domains that promises further insight into central issues in gene transmission and expression.