Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 34
Filter
Add more filters










Publication year range
1.
Cell Rep ; 43(1): 113605, 2024 01 23.
Article in English | MEDLINE | ID: mdl-38127622

ABSTRACT

Despite the symmetrical structure of nucleosomes, in vitro studies have shown that transcription proceeds with different efficiency depending on the orientation of the DNA sequence around them. However, it is unclear whether this functional asymmetry is present in vivo and whether it could regulate transcriptional directionality. Here, we report that the proximal and distal halves of nucleosomal DNA contribute differentially to nucleosome stability in the genome. In +1 nucleosomes, this asymmetry facilitates or hinders transcription depending on the orientation of its underlying DNA, and this difference is associated with an asymmetrical interaction between DNA and histones. These properties are encoded in the DNA signature of +1 nucleosomes, since its incorporation in the two orientations into downstream nucleosomes renders them asymmetrically accessible to MNase and inverts the balance between sense and antisense transcription. Altogether, our results show that nucleosomal DNA endows nucleosomes with asymmetrical properties that modulate the directionality of transcription.


Subject(s)
Histones , Nucleosomes , Histones/metabolism , DNA/metabolism , Genome , Nucleotide Motifs
2.
Cell ; 186(24): 5237-5253.e22, 2023 11 22.
Article in English | MEDLINE | ID: mdl-37944512

ABSTRACT

Here, we report the design, construction, and characterization of a tRNA neochromosome, a designer chromosome that functions as an additional, de novo counterpart to the native complement of Saccharomyces cerevisiae. Intending to address one of the central design principles of the Sc2.0 project, the ∼190-kb tRNA neochromosome houses all 275 relocated nuclear tRNA genes. To maximize stability, the design incorporates orthogonal genetic elements from non-S. cerevisiae yeast species. Furthermore, the presence of 283 rox recombination sites enables an orthogonal tRNA SCRaMbLE system. Following construction in yeast, we obtained evidence of a potent selective force, manifesting as a spontaneous doubling in cell ploidy. Furthermore, tRNA sequencing, transcriptomics, proteomics, nucleosome mapping, replication profiling, FISH, and Hi-C were undertaken to investigate questions of tRNA neochromosome behavior and function. Its construction demonstrates the remarkable tractability of the yeast model and opens up opportunities to directly test hypotheses surrounding these essential non-coding RNAs.


Subject(s)
Chromosomes, Artificial, Yeast , Genome, Fungal , Saccharomyces cerevisiae , Gene Expression Profiling , Proteomics , Saccharomyces cerevisiae/genetics , Synthetic Biology , RNA, Transfer/genetics , Chromosomes, Artificial, Yeast/genetics
3.
Genetics ; 222(1)2022 08 30.
Article in English | MEDLINE | ID: mdl-35866610

ABSTRACT

The stability of the genome is occasionally challenged by the formation of DNA-RNA hybrids and R-loops, which can be influenced by the chromatin context. This is mainly due to the fact that DNA-RNA hybrids hamper the progression of replication forks, leading to fork stalling and, ultimately, DNA breaks. Through a specific screening of chromatin modifiers performed in the yeast Saccharomyces cerevisiae, we have found that the Rtt109 histone acetyltransferase is involved in several steps of R-loop-metabolism and their associated genetic instability. On the one hand, Rtt109 prevents DNA-RNA hybridization by the acetylation of histone H3 lysines 14 and 23 and, on the other hand, it is involved in the repair of replication-born DNA breaks, such as those that can be caused by R-loops, by acetylating lysines 14 and 56. In addition, Rtt109 loss renders cells highly sensitive to replication stress in combination with R-loop-accumulating THO-complex mutants. Our data evidence that the chromatin context simultaneously influences the occurrence of DNA-RNA hybrid-associated DNA damage and its repair, adding complexity to the source of R-loop-associated genetic instability.


Subject(s)
Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae , Acetylation , Chromatin , DNA Replication , Genomic Instability , Histone Acetyltransferases/genetics , Histone Acetyltransferases/metabolism , Homeostasis , R-Loop Structures , RNA , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism
4.
Cell Rep ; 38(2): 110201, 2022 01 11.
Article in English | MEDLINE | ID: mdl-35021102

ABSTRACT

Homologous recombination is essential to maintain genome stability in response to DNA damage. Here, we have used genome-wide sequencing to quantitatively analyze at nucleotide resolution the dynamics of DNA end resection, re-synthesis, and gene conversion at a double-strand break. Resection initiates asymmetrically in an MRX-independent manner before proceeding steadily in both directions. Sgs1, Exo1, Rad51, and Srs2 differently regulate the rate and symmetry of early and late resection. Exo1 also ensures the coexistence of resection and re-synthesis, while Srs2 guarantees a constant and symmetrical DNA re-polymerization. Gene conversion is MMR independent, spans only a minor fraction of the resected region, and its unidirectionality depends on Srs2. Finally, these repair factors prevent the development of alterations remote from the DNA lesion, such as subtelomeric instability, duplication of genomic regions, and over-replication of Ty elements. Altogether, this approach allows a quantitative analysis and a direct genome-wide visualization of DNA repair by homologous recombination.


Subject(s)
Recombinational DNA Repair/genetics , Recombinational DNA Repair/physiology , DNA Breaks, Double-Stranded , DNA Damage , DNA Helicases/genetics , DNA Helicases/metabolism , DNA Repair/genetics , DNA Repair/physiology , DNA Replication , Exodeoxyribonucleases/genetics , Genome-Wide Association Study , Genomic Instability , Rad51 Recombinase/genetics , RecQ Helicases/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae Proteins/genetics , Sequence Analysis, DNA/methods
5.
Nat Struct Mol Biol ; 26(10): 970-979, 2019 10.
Article in English | MEDLINE | ID: mdl-31582854

ABSTRACT

Cohesin is a regulator of genome architecture with roles in sister chromatid cohesion and chromosome compaction. The recruitment and mobility of cohesin complexes on DNA is restricted by nucleosomes. Here, we show that the role of cohesin in chromosome organization requires the histone chaperone FACT ('facilitates chromatin transcription') in Saccharomyces cerevisiae. We find that FACT interacts directly with cohesin, and is dynamically required for its localization on chromatin. Depletion of FACT in metaphase cells prevents cohesin accumulation at pericentric regions and causes reduced binding on chromosome arms. Using the Hi-C technique, we show that cohesin-dependent TAD (topological associated domain)-like structures in G1 and metaphase chromosomes are reduced in the absence of FACT. Sister chromatid cohesion is intact in FACT-depleted cells, although chromosome segregation failure is observed. Our data show that FACT contributes to the formation of cohesin-dependent TADs, thus uncovering a new role for this complex in nuclear organization during interphase and mitotic chromosome folding.


Subject(s)
Cell Cycle Proteins/metabolism , Chromosomal Proteins, Non-Histone/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/metabolism , Transcriptional Elongation Factors/metabolism , Chromatin/metabolism , DNA-Binding Proteins/metabolism , High Mobility Group Proteins/metabolism , Protein Interaction Maps , Saccharomyces cerevisiae/cytology , Cohesins
6.
Bioinformatics ; 35(13): 2185-2192, 2019 07 01.
Article in English | MEDLINE | ID: mdl-30496344

ABSTRACT

MOTIVATION: The Burrows-Wheeler transform (BWT) is widely used for the fast alignment of high-throughput sequence data. This method also has potential applications in other areas of bioinformatics, and it can be specially useful for the fast searching of patterns on coverage data from different sources. RESULTS: We present a nucleosome pattern search method that converts levels of nucleosomal occupancy to a sequence-like format to which BWT searches can be applied. The method is embedded in a nucleosome map browser, 'Nucleosee', an interactive visual tool specifically designed to enhance BWT searches, giving them context and making them suitable for visual discourse analysis of the results. The proposed method is fast, flexible and sufficiently generic for the exploration of data in a broad and interactive way. AVAILABILITY AND IMPLEMENTATION: The proposed algorithm and visual browser are available for testing at http://cpg3.der.usal.es/nucleosee. The source code and installation packages are also available at https://github.com/rodrigoSantamaria/nucleosee. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.


Subject(s)
Genome , Nucleosomes , Algorithms , Software
7.
Methods Mol Biol ; 1766: 3-13, 2018.
Article in English | MEDLINE | ID: mdl-29605844

ABSTRACT

The discovery of CpG islands (CGIs) and the study of their structure and properties run parallel to the development of molecular biology in the last two decades of the twentieth century and to the development of high-throughput genomic technologies at the turn of the millennium. First identified as discrete G + C-rich regions of unmethylated DNA in several vertebrates, CGIs were soon found to display additional distinctive chromatin features from the rest of the genome in terms of accessibility and of the epigenetic modifications of their histones. These features, together with their colocalization with promoters and with origins of DNA replication in mammals, highlighted their relevance in the regulation of genomic processes. Recent approaches have shown with unprecedented detail the dynamics and diversity of the epigenetic landscape of CGIs during normal development and under pathological conditions. Also, comparative analyses across species have started revealing how CGIs evolve and contribute to the evolution of the vertebrate genome.


Subject(s)
CpG Islands/genetics , DNA Methylation , DNA/metabolism , Epigenesis, Genetic , Animals , Chromatin/chemistry , Chromatin/metabolism , DNA/chemistry , DNA Replication , Histones/chemistry , Histones/metabolism , Humans , Promoter Regions, Genetic/genetics , Transcription, Genetic
8.
Cold Spring Harb Protoc ; 2018(3)2018 03 01.
Article in English | MEDLINE | ID: mdl-28733396

ABSTRACT

Two-dimensional gel electrophoresis allows direct detection of DNA replication and recombination intermediates in preparations of total genomic DNA. This technique is widely used to identify replication origins in the yeast genome and is based on the different mobility in agarose gels of linear and branched DNA molecules depending on their mass and structure. During the first dimension, low-voltage and a low-percentage agarose gel favors separation of the molecules by their mass and minimizes the effect of their structure. In contrast, during the second dimension, a higher voltage, a higher percentage agarose gel, and the presence of ethidium bromide significantly delays the migration of branched structures relative to linear molecules of the same mass. This technique is appropriate for the detection of replication initiation and, therefore, an active origin of replication, within regions of approximately 2.5-5.5 kb. Consequently, it is not well suited for genome-wide replication analyses.


Subject(s)
DNA Replication , Electrophoresis, Gel, Two-Dimensional/methods , Schizosaccharomyces/genetics , DNA, Fungal/isolation & purification
9.
Cold Spring Harb Protoc ; 2018(4)2018 04 02.
Article in English | MEDLINE | ID: mdl-28733402

ABSTRACT

The fission yeast Schizosaccharomyces pombe is an excellent model organism to study DNA metabolism, in which the DNA replication and repair mechanisms are evolutionarily conserved. In this introduction we describe a range of methods commonly used to study aspects of DNA metabolism in fission yeast, focusing on approaches used for the analysis of genome stability, DNA replication, and DNA repair. We describe the use of a minichromosome, Ch16, for monitoring different aspects of genome stability. We introduce two-dimensional gel electrophoresis and immunofluorescent visualization of combed DNA molecules for the analysis of DNA replication. Further, we introduce a pulsed field gel electrophoresis (PFGE) assay to physically monitor chromosome integrity, which can be used in conjunction with a DNA double-strand break (DSB) repair assay to genetically quantitate different DSB repair and misrepair outcomes, including gross chromosomal rearrangements, in fission yeast.


Subject(s)
Biochemistry/methods , DNA, Fungal/metabolism , Schizosaccharomyces/metabolism , DNA Repair , DNA Replication , Genomic Instability , Schizosaccharomyces/genetics
10.
11.
Nucleus ; 8(5): 469-474, 2017 09 03.
Article in English | MEDLINE | ID: mdl-28635365

ABSTRACT

Nucleosomes are the basic units of chromatin. They compact the genome inside the nucleus and regulate the access of proteins to DNA. In the yeast genome, most nucleosomes occupy well-defined positions, which are maintained under many different physiological situations and genetic backgrounds. Although several short sequence elements have been described that favor or reduce the affinity between histones and DNA, the extent to which the DNA sequence affects nucleosome positioning in the genomic context remains unclear. Recent analyses indicate that the base composition pattern of mononucleosomal DNA differs among species, and that the same sequence elements have a different impact on nucleosome positioning in different genomes despite the high level of phylogenetic conservation of histones. These studies have also shown that the DNA sequence contributes to nucleosome positioning to the point that it is possible to design synthetic DNA molecules capable of generating regular and species-specific nucleosomal patterns in vivo.


Subject(s)
DNA/chemistry , Nucleosomes/metabolism , Base Composition , DNA/metabolism , Saccharomyces cerevisiae/cytology , Saccharomyces cerevisiae/genetics , Schizosaccharomyces/cytology , Schizosaccharomyces/genetics
12.
Curr Genet ; 63(2): 187-193, 2017 May.
Article in English | MEDLINE | ID: mdl-27558480

ABSTRACT

The occupancy of nucleosomes governs access to the eukaryotic genomes and results from a combination of biophysical features and the effect of ATP-dependent remodelling complexes. Most promoter regions show a conserved pattern characterized by a nucleosome-depleted region (NDR) flanked by nucleosomal arrays. The conserved RSC remodeler was reported to be critical to establish NDR in vivo in budding yeast but other evidences suggested that this activity may not be conserved in fission yeast. By reanalysing and expanding previously published data, we propose that NDR formation requires, at least partially, RSC in both yeast species. We also discuss the most prominent biological role of RSC and the possibility that non-essential subunits do not define alternate versions of the complex.


Subject(s)
Adenosine Triphosphatases/genetics , Chromatin Assembly and Disassembly/genetics , Nucleosomes/genetics , Saccharomyces cerevisiae Proteins/genetics , Schizosaccharomyces pombe Proteins/genetics , Adenosine Triphosphatases/metabolism , Gene Expression Profiling , Gene Expression Regulation, Fungal , Saccharomyces cerevisiae Proteins/metabolism , Saccharomycetales/genetics , Saccharomycetales/metabolism , Schizosaccharomyces/genetics , Schizosaccharomyces/metabolism , Schizosaccharomyces pombe Proteins/metabolism , Species Specificity
13.
Genome Res ; 26(11): 1532-1543, 2016 11.
Article in English | MEDLINE | ID: mdl-27662899

ABSTRACT

In the yeast genome, a large proportion of nucleosomes occupy well-defined and stable positions. While the contribution of chromatin remodelers and DNA binding proteins to maintain this organization is well established, the relevance of the DNA sequence to nucleosome positioning in the genome remains controversial. Through quantitative analysis of nucleosome positioning, we show that sequence changes distort the nucleosomal pattern at the level of individual nucleosomes in three species of Schizosaccharomyces and in Saccharomyces cerevisiae This effect is equally detected in transcribed and nontranscribed regions, suggesting the existence of sequence elements that contribute to positioning. To identify such elements, we incorporated information from nucleosomal signatures into artificial synthetic DNA molecules and found that they generated regular nucleosomal arrays indistinguishable from those of endogenous sequences. Strikingly, this information is species-specific and can be combined with coding information through the use of synonymous codons such that genes from one species can be engineered to adopt the nucleosomal organization of another. These findings open the possibility of designing coding and noncoding DNA molecules capable of directing their own nucleosomal organization.


Subject(s)
Genome, Fungal , Nucleosomes/genetics , Open Reading Frames , Regulatory Sequences, Nucleic Acid , Saccharomyces cerevisiae/genetics , Schizosaccharomyces/genetics
14.
Elife ; 52016 05 12.
Article in English | MEDLINE | ID: mdl-27171419

ABSTRACT

In fission yeast, the ste11 gene encodes the master regulator initiating the switch from vegetative growth to gametogenesis. In a previous paper, we showed that the methylation of H3K4 and consequent promoter nucleosome deacetylation repress ste11 induction and cell differentiation (Materne et al., 2015) but the regulatory steps remain poorly understood. Here we report a genetic screen that highlighted H2B deubiquitylation and the RSC remodeling complex as activators of ste11 expression. Mechanistic analyses revealed more complex, opposite roles of H2Bubi at the promoter where it represses expression, and over the transcribed region where it sustains it. By promoting H3K4 methylation at the promoter, H2Bubi initiates the deacetylation process, which decreases chromatin remodeling by RSC. Upon induction, this process is reversed and efficient NDR (nucleosome depleted region) formation leads to high expression. Therefore, H2Bubi represses gametogenesis by opposing the recruitment of RSC at the promoter of the master regulator ste11 gene.


Subject(s)
Chromatin Assembly and Disassembly , Gene Expression Regulation, Fungal , Histones/metabolism , MAP Kinase Kinase Kinases/metabolism , Schizosaccharomyces/cytology , Schizosaccharomyces/genetics , Transcription Factors/antagonists & inhibitors , Ubiquitination
15.
Bioessays ; 37(10): 1067-73, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26293347

ABSTRACT

The dynamics of eukaryotic DNA polymerases has been difficult to establish because of the difficulty of tracking them along the chromosomes during DNA replication. Recent work has addressed this problem in the yeasts Schizosaccharomyces pombe and Saccharomyces cerevisiae through the engineering of replicative polymerases to render them prone to incorporating ribonucleotides at high rates. Their use as tracers of the passage of each polymerase has provided a picture of unprecedented resolution of the organization of replicons and replication origins in the two yeasts and has uncovered important differences between them. Additional studies have found an overlapping distribution of DNA polymorphisms and the junctions of Okazaki fragments along mononucleosomal DNA. This sequence instability is caused by the premature release of polymerase δ and the retention of non proof-read DNA tracts replicated by polymerase α. The possible implementation of these new experimental approaches in multicellular organisms opens the door to the analysis of replication dynamics under a broad range of genetic backgrounds and physiological or pathological conditions.


Subject(s)
DNA Replication , DNA-Directed DNA Polymerase/metabolism , Saccharomycetales/genetics , Schizosaccharomyces/genetics , DNA , DNA, Fungal/metabolism , DNA-Directed DNA Polymerase/analysis , Genome, Fungal , Genomic Instability , Replication Origin , Saccharomyces cerevisiae/genetics
16.
Elife ; 4: e09008, 2015 Jun 22.
Article in English | MEDLINE | ID: mdl-26098123

ABSTRACT

The phosphorylation of the RNA polymerase II C-terminal domain (CTD) plays a key role in delineating transcribed regions within chromatin by recruiting histone methylases and deacetylases. Using genome-wide nucleosome mapping, we show that CTD S2 phosphorylation controls nucleosome dynamics in the promoter of a subset of 324 genes, including the regulators of cell differentiation ste11 and metabolic adaptation inv1. Mechanistic studies on these genes indicate that during gene activation a local increase of phospho-S2 CTD nearby the promoter impairs the phospho-S5 CTD-dependent recruitment of Set1 and the subsequent recruitment of specific HDACs, which leads to nucleosome depletion and efficient transcription. The early increase of phospho-S2 results from the phosphorylation of the CTD S2 kinase Lsk1 by MAP kinase in response to cellular signalling. The artificial tethering of the Lsk1 kinase at the ste11 promoter is sufficient to activate transcription. Therefore, signalling through the CTD code regulates promoter nucleosomes dynamics.


Subject(s)
Nucleosomes/metabolism , Protein Processing, Post-Translational , RNA Polymerase II/metabolism , Gene Expression Profiling , Gene Expression Regulation, Fungal , Molecular Sequence Data , Phosphorylation , Promoter Regions, Genetic , Schizosaccharomyces/enzymology , Schizosaccharomyces/genetics , Schizosaccharomyces/metabolism , Sequence Analysis, DNA , Signal Transduction , Transcription, Genetic , Transcriptional Activation
17.
Open Biol ; 5(4): 140218, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25854683

ABSTRACT

Nucleosomes are the basic structural units of chromatin. Most of the yeast genome is organized in a pattern of positioned nucleosomes that is stably maintained under a wide range of physiological conditions. In this work, we have searched for sequence determinants associated with positioned nucleosomes in four species of fission and budding yeasts. We show that mononucleosomal DNA follows a highly structured base composition pattern, which differs among species despite the high degree of histone conservation. These nucleosomal signatures are present in transcribed and non-transcribed regions across the genome. In the case of open reading frames, they correctly predict the relative distribution of codons on mononucleosomal DNA, and they also determine a periodicity in the average distribution of amino acids along the proteins. These results establish a direct and species-specific connection between the position of each codon around the histone octamer and protein composition.


Subject(s)
Amino Acids/genetics , Fungal Proteins/genetics , Gene Expression Regulation, Fungal , Nucleosomes/genetics , Transcriptome , Amino Acids/metabolism , DNA, Fungal/genetics , Fungal Proteins/metabolism , Genome, Fungal/genetics , Nucleosomes/metabolism , Open Reading Frames/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Schizosaccharomyces/classification , Schizosaccharomyces/genetics , Schizosaccharomyces/metabolism , Species Specificity
18.
Brief Bioinform ; 16(4): 576-87, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25296770

ABSTRACT

Nucleosomes contribute to compacting the genome into the nucleus and regulate the physical access of regulatory proteins to DNA either directly or through the epigenetic modifications of the histone tails. Precise mapping of nucleosome positioning across the genome is, therefore, essential to understanding the genome regulation. In recent years, several experimental protocols have been developed for this purpose that include the enzymatic digestion, chemical cleavage or immunoprecipitation of chromatin followed by next-generation sequencing of the resulting DNA fragments. Here, we compare the performance and resolution of these methods from the initial biochemical steps through the alignment of the millions of short-sequence reads to a reference genome to the final computational analysis to generate genome-wide maps of nucleosome occupancy. Because of the lack of a unified protocol to process data sets obtained through the different approaches, we have developed a new computational tool (NUCwave), which facilitates their analysis, comparison and assessment and will enable researchers to choose the most suitable method for any particular purpose. NUCwave is freely available at http://nucleosome.usal.es/nucwave along with a step-by-step protocol for its use.


Subject(s)
Genome-Wide Association Study , Nucleosomes/genetics , Chromatin Immunoprecipitation , Humans , Sequence Alignment
19.
Cell ; 159(3): 572-83, 2014 Oct 23.
Article in English | MEDLINE | ID: mdl-25417108

ABSTRACT

Nuclear RNAi is an important regulator of transcription and epigenetic modification, but the underlying mechanisms remain elusive. Using a genome-wide approach in the fission yeast S. pombe, we have found that Dcr1, but not other components of the canonical RNAi pathway, promotes the release of Pol II from the 3? end of highly transcribed genes, and, surprisingly, from antisense transcription of rRNA and tRNA genes, which are normally transcribed by Pol I and Pol III. These Dcr1-terminated loci correspond to sites of replication stress and DNA damage, likely resulting from transcription-replication collisions. At the rDNA loci, release of Pol II facilitates DNA replication and prevents homologous recombination, which would otherwise lead to loss of rDNA repeats especially during meiosis. Our results reveal a novel role for Dcr1-mediated transcription termination in genome maintenance and may account for widespread regulation of genome stability by nuclear RNAi in higher eukaryotes.


Subject(s)
DNA Replication , Endoribonucleases/metabolism , Genomic Instability , Schizosaccharomyces pombe Proteins/metabolism , Schizosaccharomyces/metabolism , Transcription Termination, Genetic , DNA, Antisense/genetics , DNA, Antisense/metabolism , RNA Interference , RNA Polymerase II/metabolism , Schizosaccharomyces/enzymology , Transcription, Genetic
20.
BMC Genomics ; 15: 791, 2014 Sep 13.
Article in English | MEDLINE | ID: mdl-25218085

ABSTRACT

BACKGROUND: Eukaryotic genomes are replicated during S phase according to a temporal program. Several determinants control the timing of origin firing, including the chromatin environment and epigenetic modifications. However, how chromatin structure influences the timing of the activation of specific origins is still poorly understood. RESULTS: By performing high-resolution analysis of genome-wide nucleosome positioning we have identified different chromatin architectures at early and late replication origins. These different patterns are already established in G1 and are tightly correlated with the organization of adjacent transcription units. Moreover, specific early and late nucleosomal patterns are fixed robustly, even in rpd3 mutants in which histone acetylation and origin timing have been significantly altered. Nevertheless, higher histone acetylation levels correlate with the local modulation of chromatin structure, leading to increased origin accessibility. In addition, we conducted parallel analyses of replication and nucleosome dynamics that revealed that chromatin structure at origins is modulated during origin activation. CONCLUSIONS: Our results show that early and late replication origins present distinctive nucleosomal configurations, which are preferentially associated to different genomic regions. Our data also reveal that origin structure is dynamic and can be locally modulated by histone deacetylation, as well as by origin activation. These data offer novel insight into the contribution of chromatin structure to origin selection and firing in budding yeast.


Subject(s)
DNA Replication Timing , DNA, Fungal/genetics , Histone Deacetylases/genetics , Nucleosomes/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae/cytology , Saccharomyces cerevisiae/genetics , Acetylation , Chromatin Assembly and Disassembly , Histones/metabolism , Mutation , Replication Origin , Saccharomyces cerevisiae Proteins/metabolism
SELECTION OF CITATIONS
SEARCH DETAIL
...