Evolution of the histone variant H2A.W reconstituted in fission yeast [ChIP-seq]
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ABSTRACT: Diversification of histone variants is marked by the acquisition of distinct motifs and features through convergent evolution. H2A variants tend to be associated with defined domains of the genome. Specific features distinguish H2A variants in eukaryotes but whether evolution of these features predated the evolution of deposition mechanisms or vice-versa has remained unclear.In flowering plants, the variant H2A.W is tightly associated with heterochromatin. H2A.W evolved in land plants through acquisition of an extended C-terminal tail enriched with basic residues and a KSPK motif. Here, we used a synthetic approach in fission yeast, which lacks H2A.W and its dedicated deposition mechanism, to recapitulate the evolutionary steps that led to H2A.W and to assess the impact of the KSPK motif on heterochromatin composition and its properties. In conclusion, the acquisition of the KSPK motif in yeast promotes chromatin properties that are comparable to the properties and function of H2A.W in plant heterochromatin. Hence, the KSPK motif could have been selected before the evolution of direct heterochromatin deposition mechanisms. We propose that the acquisition of functional histone variant motifs can confer properties which affect only specific chromatin states, thereby driving the evolution of specific deposition mechanisms.
Project description:Diversification of histone variants is marked by the acquisition of distinct motifs and features through convergent evolution. H2A variants tend to be associated with defined domains of the genome. Specific features distinguish H2A variants in eukaryotes but whether evolution of these features predated the evolution of deposition mechanisms or vice-versa has remained unclear.In flowering plants, the variant H2A.W is tightly associated with heterochromatin. H2A.W evolved in land plants through acquisition of an extended C-terminal tail enriched with basic residues and a KSPK motif. Here, we used a synthetic approach in fission yeast, which lacks H2A.W and its dedicated deposition mechanism, to recapitulate the evolutionary steps that led to H2A.W and to assess the impact of the KSPK motif on heterochromatin composition and its properties. In conclusion, the acquisition of the KSPK motif in yeast promotes chromatin properties that are comparable to the properties and function of H2A.W in plant heterochromatin. Hence, the KSPK motif could have been selected before the evolution of direct heterochromatin deposition mechanisms. We propose that the acquisition of functional histone variant motifs can confer properties which affect only specific chromatin states, thereby driving the evolution of specific deposition mechanisms.
Project description:Diversification of histone variants is marked by the acquisition of distinct motifs and features through convergent evolution. H2A variants tend to be associated with defined domains of the genome. Specific features distinguish H2A variants in eukaryotes but whether evolution of these features predated the evolution of deposition mechanisms or vice-versa has remained unclear.In flowering plants, the variant H2A.W is tightly associated with heterochromatin. H2A.W evolved in land plants through acquisition of an extended C-terminal tail enriched with basic residues and a KSPK motif. Here, we used a synthetic approach in fission yeast, which lacks H2A.W and its dedicated deposition mechanism, to recapitulate the evolutionary steps that led to H2A.W and to assess the impact of the KSPK motif on heterochromatin composition and its properties. In conclusion, the acquisition of the KSPK motif in yeast promotes chromatin properties that are comparable to the properties and function of H2A.W in plant heterochromatin. Hence, the KSPK motif could have been selected before the evolution of direct heterochromatin deposition mechanisms. We propose that the acquisition of functional histone variant motifs can confer properties which affect only specific chromatin states, thereby driving the evolution of specific deposition mechanisms.
Project description:Histone variants play crucial roles in gene expression, genome integrity and chromosome segregation. However, to what extent histone variants control chromatin architecture remains largely unknown. Here, we show that the previously uncharacterized histone variant H2A.W plays a crucial role in condensation of heterochromatin. Genome-wide profiling of all four types of H2A variants in Arabidopsis shows that H2A.W specifically associates with heterochromatin. H2A.W recruitment is independent of heterochromatic marks H3K9me2 and DNA methylation. Genetic interactions show that H2A.W acts in synergy with CMT3 mediated methylation to maintain genome integrity. In vitro, H2A.W enhances chromatin condensation through a higher propensity to make fiber-to-fiber interactions via its conserved C-terminal motif. In vivo, elimination of H2A.W causes decondensation of heterochromatin and conversely, ectopic expression of H2A.W promotes heterochromatin condensation. These results demonstrate that H2A.W plays critical roles in heterochromatin by promoting higher order chromatin condensation. Since similar H2A.W C-terminal motifs are present in other variant found in mammals and other organisms our findings impact our understanding of heterochromatin condensation in a wide variety of eukaryotic organisms. Two mRNA-seq samples, two bisulfite-seq samples, six ChIP-seq samples.
Project description:Histone variants play crucial roles in gene expression, genome integrity and chromosome segregation. However, to what extent histone variants control chromatin architecture remains largely unknown. Here, we show that the previously uncharacterized histone variant H2A.W plays a crucial role in condensation of heterochromatin. Genome-wide profiling of all four types of H2A variants in Arabidopsis shows that H2A.W specifically associates with heterochromatin. H2A.W recruitment is independent of heterochromatic marks H3K9me2 and DNA methylation. Genetic interactions show that H2A.W acts in synergy with CMT3 mediated methylation to maintain genome integrity. In vitro, H2A.W enhances chromatin condensation through a higher propensity to make fiber-to-fiber interactions via its conserved C-terminal motif. In vivo, elimination of H2A.W causes decondensation of heterochromatin and conversely, ectopic expression of H2A.W promotes heterochromatin condensation. These results demonstrate that H2A.W plays critical roles in heterochromatin by promoting higher order chromatin condensation. Since similar H2A.W C-terminal motifs are present in other variant found in mammals and other organisms our findings impact our understanding of heterochromatin condensation in a wide variety of eukaryotic organisms.
Project description:Silencing of transposons by the chromatin remodeler DDM1 is mediated by the deposition of heterochromatic H2A variants. Transposon mobility and silencing participates in genome evolution but also threaten genome integrity. DECREASED DNA METHYLATION 1 (DDM1) belongs to a conserved family of chromatin remodelers that are required to silence transposons, yet the underlying molecular mechanism has remained unknown. Here we show that DDM1 binds the histone variant H2A.W through two conserved domains that are required to deposit H2A.W and maintain transposon silencing. The mechanism of transcriptional silencing described here is likely shared among chromatin remodelers of the DDM1 family and heterochromatic H2A variants that have evolved in mammals.
Project description:Silencing of transposons by the chromatin remodeler DDM1 is mediated by the deposition of heterochromatic H2A variants. Transposon mobility and silencing participates in genome evolution but also threaten genome integrity. DECREASED DNA METHYLATION 1 (DDM1) belongs to a conserved family of chromatin remodelers that are required to silence transposons, yet the underlying molecular mechanism has remained unknown. Here we show that DDM1 binds the histone variant H2A.W through two conserved domains that are required to deposit H2A.W and maintain transposon silencing. The mechanism of transcriptional silencing described here is likely shared among chromatin remodelers of the DDM1 family and heterochromatic H2A variants that have evolved in mammals.
Project description:Silencing of transposons by the chromatin remodeler DDM1 is mediated by the deposition of heterochromatic H2A variants. Transposon mobility and silencing participates in genome evolution but also threaten genome integrity. DECREASED DNA METHYLATION 1 (DDM1) belongs to a conserved family of chromatin remodelers that are required to silence transposons, yet the underlying molecular mechanism has remained unknown. Here we show that DDM1 binds the histone variant H2A.W through two conserved domains that are required to deposit H2A.W and maintain transposon silencing. The mechanism of transcriptional silencing described here is likely shared among chromatin remodelers of the DDM1 family and heterochromatic H2A variants that have evolved in mammals.
Project description:In flowering plants, heterochromatin is demarcated by the histone variant H2A.W, elevated levels of the linker histone H1, and specific epigenetic modifications, such as high levels of DNA methylation at both CG and non-CG sites. How H2A.W regulates heterochromatin organization and interacts with other heterochromatic features is unclear. Here, we create an h2a.w null mutant via CRISPR-Cas9, h2a.w-2, to analyze the in vivo function of H2A.W. We find that H2A.W antagonizes deposition of H1 at heterochromatin and that non-CG methylation and accessibility are moderately decreased in h2a.w-2 heterochromatin. Compared to H1 loss alone, combined loss of H1 and H2A.W greatly increases accessibility and facilitates non-CG DNA methylation in heterochromatin, suggesting co-regulation of heterochromatic features by H2A.W and H1. Our results suggest that H2A.W helps maintain optimal heterochromatin accessibility and DNA methylation by promoting chromatin compaction together with H1, while also inhibiting excessive H1 incorporation.
Project description:Programmed constitutive heterochromatin silencing is essential for eukaryotic genome regulation, yet the initial step of this process is ambiguous. A large proportion of R-loops (RNA:DNA hybrids) had been unexpectedly identified within Arabidopsis pericentromeric heterochromatin with unknown functions. Through a genome-wide R-loop profiling screen, we find DDM1 (Decrease in DNA Methylation 1) is the primary restrictor of pericentromeric R-loops via its RNA:DNA helicase activity. Low levels of pericentromeric R-loops resolved by DDM1 co-transcriptionally can prime constitutive heterochromatin silencing. Furthermore, we demonstrate that DDM1 physically excludes histone H2A variant H2A.Z, and promotes H2A.W deposition for faithful heterochromatin initiation soon after R-loop clearance. The dual functions of DDM1 in R-loop resolution and H2A.Z eviction are essential for sperm nuclei structure maintenance in mature pollen. Our work unravels the co-transcriptional R-loop resolution coupled with accurate H2A variants deposition is the primary step of constitutive heterochromatin silencing in Arabidopsis, which might be conserved across eukaryotes.
Project description:Programmed constitutive heterochromatin silencing is essential for eukaryotic genome regulation, yet the initial step of this process is ambiguous. A large proportion of R-loops (RNA:DNA hybrids) had been unexpectedly identified within Arabidopsis pericentromeric heterochromatin with unknown functions. Through a genome-wide R-loop profiling screen, we find DDM1 (Decrease in DNA Methylation 1) is the primary restrictor of pericentromeric R-loops via its RNA:DNA helicase activity. Low levels of pericentromeric R-loops resolved by DDM1 co-transcriptionally can prime constitutive heterochromatin silencing. Furthermore, we demonstrate that DDM1 physically excludes histone H2A variant H2A.Z, and promotes H2A.W deposition for faithful heterochromatin initiation soon after R-loop clearance. The dual functions of DDM1 in R-loop resolution and H2A.Z eviction are essential for sperm nuclei structure maintenance in mature pollen. Our work unravels the co-transcriptional R-loop resolution coupled with accurate H2A variants deposition is the primary step of constitutive heterochromatin silencing in Arabidopsis, which might be conserved across eukaryotes.