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Szymanska-Lejman, M.

Publications and source records attributed to Szymanska-Lejman, M..

4 recordsLinked to original sources

Chromatin crosstalk between HDA19 and NuA4 sets thresholds for stress gene activation in Arabidopsis

Histone acetylation by the plant NuA4 complex promotes high expression of growth-related genes while preventing gene body H2A.Z depletion and spurious activation of stress-responsive loci. How this NuA4-dependent chromatin state is modulated by histone deacetylases (HDACs) in plants remains unclear. Here, we investigate the interplay between NuA4 and HDACs in Arabidopsis by generating a collection of HDAC loss-of-function mutants in NuA4-proficient and NuA4-deficient backgrounds. Loss of individual HDACs did not rescue the severe growth defects of the NuA4(-) mutant; instead, most combinations further aggravated the phenotype, indicating that HDACs predominantly support rather than counteract NuA4 function. Focusing on the hda19-5Atepl1ballele, we show that loss of HDA19 causes strong developmental defects and constitutive activation of biotic stress-responsive genes, but not canonical heat-response genes. 3' RNA-seq and ChIP-seq reveal that upregulated genes in hda19-5Atepl1bdisplay increased H3K9ac and H2A.Zac with largely unchanged gene-body H2A.Z, and substantially overlap with genes induced in the NuA4-null Atepl1-2 mutant and in wild-type plants exposed to elevated temperature. Integration of our datasets with published HDA19 ChIP-seq maps shows that H2A.Zac is selectively increased at HDA19-bound H2A.Z peaks in hda19-5Atepl1b, consistent with a direct role for HDA19 in H2A.Z deacetylation. However, elevated H2A.Zac is neither necessary nor sufficient for transcriptional activation, whereas changes in H3K9ac correlate strongly with gene induction. We propose that NuA4 and HDA19 cooperate to tune chromatin at stress-related loci, with H3K9 acetylation as the primary driver of transcription and H2A.Z acetylation acting as a modulatory mark that shapes stress gene responsiveness.

plant biology↗

Modifying meiotic recombination by targeting chromatin modifiers to crossover hotspots in Arabidopsis

The impact of specific chromatin modifications on meiotic crossover frequency is usually inferred from correlative studies, leaving open the question of causality. To address this, we used a dCas9-based system to target histone H3 methylation modifiers to defined genomic loci. Targeting methyltransferases responsible for H3K9me3 and H3K27me3 had little effect on recombination at selected hotspots, whereas targeting H3K4me3- and H3K36me3-associated enzymes often silenced the endogenous genes. The strongest effect was observed with the demethylase JMJ14, which reduced local H3K4me3 levels and decreased crossover frequency within the targeted interval. This was accompanied by reduced transcription of a long non-coding RNA (lncRNA) located at the hotspot and altered crossover topology. Suppressed recombination was also seen at neighbouring, untargeted hotspots. Conversely, directing the transcriptional activator VP64 to the same region elevated lncRNA expression, increased crossover frequency, and raised H3K4me3 levels. Our results reveal a causal relationship between H3K4me3, transcription, and local crossover activity, demonstrating that H3K4me3 levels are tightly associated with both transcriptional output and recombination frequency at specific genomic sites.

genetics↗

MSH2 stimulates interfering and inhibits non-interfering crossovers in response to genetic polymorphism

In Arabidopsis, local sequence polymorphism between homologs can stimulate double-strand break (DSB) repair via crossover formation in a MSH2-dependent manner. To understand how MSH2 regulates crossovers formed by the independent interfering and non-interfering pathways, we combine mutants that elevate non-interfering crossovers with msh2. We demonstrate that MSH2 blocks non-interfering crossovers at polymorphic loci, which is the opposite effect to interfering crossovers. We also observe MSH2-independent crossover inhibition at highly polymorphic sites. We measure recombination along the chromosome arms, in lines differing in the heterozygosity pattern, and observe a dramatic crossover increase at the boundaries between heterozygous and homozygous regions, which is MSH2-dependent. Together, we show that MSH2 is a master regulator of meiotic DSB repair, with antagonistic effects on interfering and non-interfering crossovers, that shapes the crossover landscape in relation to interhomolog polymorphism.

genetics↗

NuA4-dependent H4 acetylation is adopted to specifically control photosynthesis-related genes

NuA4, an essential histone acetyltransferase complex, is required for efficient transcription in eukaryotes. Using genome editing, genomic approaches and biochemical assays, we characterized plant homologues of two key components of this complex, EPL1 and EAF1 in Arabidopsis thaliana. Surprisingly, we found that loss of AtEPL1, which is necessary for enzymatic activity of NuA4, is not lethal. Contrary to yeast, mutants lacking AtEAF1, responsible for complex targeting, display severe pleiotropic phenotype which copies that of Atepl1. Atepl1 and Ateaf1 mutants grow slowly, contain reduced chlorophyll levels and small chloroplasts. We provide evidence that these alterations are not caused by disturbance of GLK transcription factors, the major regulators of chloroplast development. Using ChIP-seq we show that H4 acetylation levels are dramatically reduced in the chromatin of the Atepl1 mutant, while H3 acetylation remains mostly unchanged. We use our data to define NuA4-dependent genes and show that chloroplast-related genes are significantly overrepresented in this group, consistent with the pale-green phenotypes of the mutants. We propose that NuA4 was adopted in plants to control nuclear-encoded photosynthesis genes. SignificancePhotosynthesis depends on chloroplast proteins, most of which are nucleus-encoded and thus subject to control mechanisms common across eukaryotes. Here we show that NuA4, an evolutionary conserved transcriptional coactivator, is necessary for proper development of photosynthetic apparatus. Surprisingly, in contrast to yeast and metazoans, plants engineered to lack core NuA4 subunits are capable of vegetative development despite dramatic genome-wide loss of NuA4-dependent H4K5 acetylation. This chromatin perturbation seems to directly affect 350 genes which, in addition to reduced H4K5ac levels, display decreased transcript levels but no loss of transcription-related H3K9ac. A significant proportion of these genes are related to chloroplast function, particularly to translation, an intriguing parallel to the yeast NuA4s role in transcription of ribosome biogenesis-related genes.

molecular biology↗