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Histone H1 prevents non-CG methylation-mediated small RNA biogenesis in Arabidopsis heterochromatin

ABSTRACT: Histone H1 prevents non-CG methylation-mediated small RNA biogenesis in Arabidopsis heterochromatin

PROVIDER: PRJNA745095 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15082700_1.fastq.gz	Fastqsanger.gz
	SRR15082700_2.fastq.gz	Fastqsanger.gz
	SRR15082701_1.fastq.gz	Fastqsanger.gz
	SRR15082701_2.fastq.gz	Fastqsanger.gz
	SRR15082702_1.fastq.gz	Fastqsanger.gz

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Project description:Silencing pathways prevent transposable element (TE) proliferation and help to maintain genome integrity through cell division. Silenced genomic regions can be classified as either euchromatic or heterochromatic, and are targeted by genetically separable epigenetic pathways. In plants, the RNA-directed DNA methylation (RdDM) pathway targets mostly euchromatic regions, while CMT methyltransferases are mainly associated with heterochromatin. However, many epigenetic features - including DNA methylation patterning - are largely indistinguishable between these regions, so how the functional separation is maintained is unclear. The linker histone H1 is preferentially localised to heterochromatin and has been proposed to restrict RdDM from encroachment. To test this hypothesis, we followed RdDM genomic localisation in an H1 mutant by performing ChIP-seq on the largest subunit, NRPE1, of the central RdDM polymerase (Pol V). Loss of H1 resulted in heterochromatic TE enrichment of NPRE1. Increased NRPE1 binding was associated with increased chromatin accessibility in h1, suggesting that H1 restricts NRPE1 occupancy by compacting chromatin. However, RdDM occupancy did not impact H1 localization, demonstrating that H1 hierarchically restricts RdDM positioning. H1 mutants experience major symmetric (CG and CHG) DNA methylation gains, and by generating an h1/nrpe1 double mutant, we demonstrate these gains are largely independent of RdDM. However, loss of NRPE1 occupancy from a subset of euchromatic regions in h1 experienced corresponding loss of methylation in all sequence contexts, while at ectopically bound heterochromatic loci, NRPE1 deposited methylation specifically in the CHH context. Additionally, we found that H1 restricts the occupancy of the methylation reader and activator complex component, SUVH1, indicating that H1’s regulatory control of methylation pathways is not limited to RdDM. Together, the results support a model whereby H1 helps maintain the exclusivity of heterochromatin by preventing encroachment from other competing pathways.

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Histone H1 prevents non-CG methylation-mediated small RNA biogenesis in Arabidopsis heterochromatin

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