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PNET-seq in Arabidopsis after flavopiridol (FLA) treatment

ABSTRACT: Eukaryotic euchromatin is the less compact chromatin and modified by many histone modifications such as H3 Lysine 36 methylation (H3K36me). Here we report a new chromatin state, Transcription Resistive, which is differentiated from activation and silencing. Transcription resistive is stamped by H3K36me with almost undetectable transcription activity but open chromatin state, and occupies most documented plant essential genes. Mutating SDG8, previously known as the major H3K36 methyltransferase in Arabidopsis, surprisingly elevates 78.7% H3K36me3 marked resistive loci, which count for 39.4% of the coding genome. Genetically, SDG8 prevents H3K36me activity of SDG4 at short and intron-less genes to secure plant fertility, while SDG8 collaborates with other H3K36me methyltransferases on long and intron-rich genes. Together, our results reveal SDG8 is the primary sensor to suppress excessive H3K36me, and uncovered the resistive is the conserved H3K36me-stamped novel transcription state in plants, highlighting the regulatory diversities and biological significances of H3K36 methylation in eukaryotes.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE281306 | GEO | 2025/02/08

REPOSITORIES: GEO

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Project description:Bioinformatics powered correlative analysis of epigenomic patterns is an effective method to help derive biological hypotheses that can be tested genetically or biochemically. To accommodate the variety and complexity of epigenomic and transcriptomic patterns, ANchored COrrelative Patterns (ANCORP) was developed as a platform to integrate and intuitively visualize a large number of genome-wide profiles. With global profiles of 9 histone modifications mapped by ChIP-seq and a strand-specific RNA-seq dataset, we have applied the ANCORP-genetics pipeline for hypothesis building and testing in order to understand how global transcription may be regulated by epigenetic pathways such as histone modifications. It was found that intragenic antisense RNAs were depleted from genes with strong gene-body H3K36me2 mark and cytosine methylation enrichments but were significantly overrepresented in H3K4me3/H3K27me3 bivalent genes. Moreover, gene body H3K36me2 and DNA methylation anti-correlated with multiple active chromatin marks including H3K4me2/3, H3K9Ac and H3K18Ac. These observations lead us to hypothesize that H3K36me2 and DNA methylation may synergistically repress active chromatin marks in gene bodies and subsequently inhibit transcription of the antisense strand. Mutant analyses revealed that Polymerase Associated Factors (PAF) may be universally required for modulating NAT abundance whereas the role for the 5mC and H3K36me marks are more locus specific. H3K36me and PAF may either repress or permit the accumulation NATs depending on the chromatin state context in a particular transcription unit. Interestingly, the activation of antisense RNA in sdg8-2 or elf8-1 mutants does not associate with any increase of histone marks in gene bodies that are known to correlate with gene activation. Our results suggest that ANCORP-genetics is an effective approach to uncover epigenetic regulatory mechanisms by leveraging on the rapid advances in sequencing technologies and the resultant wealth of genome-wide information.

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PNET-seq in Arabidopsis after flavopiridol (FLA) treatment

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