NADs identification under heat-stress
Ontology highlight
ABSTRACT:
PROVIDER: PRJEB36061 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR14364900.fastq.gz | Fastqsanger.gz | |||
| ERR14364901.fastq.gz | Fastqsanger.gz | |||
| ERR14364902.fastq.gz | Fastqsanger.gz | |||
| ERR14364903.fastq.gz | Fastqsanger.gz | |||
| ERR14364904.fastq.gz | Fastqsanger.gz |
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Project description:Repressive chromatin domains often localize to the nuclear lamina or nucleolus. Although nucleolar-associated domains (NADs) have recently been mapped, their mechanisms of nucleolar association and functional significance remain unclear. Here, we show that nucleophosmin (NPM1), a factor located in the granular component of the nucleolus, mediates NAD association in mouse embryonic stem cells. NPM1 binds NADs, interacts with the histone methyltransferase G9a (EHMT2), and is required for establishing H3K9me2 at NADs. Loss of NPM1 or expression of a DNA-binding-deficient mutant disrupts NAD-nucleolus association and reduces H3K9me2 specifically at NADs. G9a is dispensable for NAD-nucleolus contacts, indicating that H3K9me2 is acquired after NADs associate with NPM1 at nucleoli. These findings reveal mechanistic insights into how genomic domains associate with nucleoli and form repressive chromatin and indicate that the nucleolus not only serves as scaffold for positioning repressive domains but also plays a direct role in establishing their repressive chromatin states.
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Project description:Heat stress (HS) impacts the nuclear proteome and, subsequently, protein activities in the nucleoplasm and/or different nuclear compartments. In Arabidopsis thaliana, a short exposure to 37°C leads to the loss of the standard tripartite architecture of the nucleolus, the most prominent nuclear substructure, and, consequently, affects the assembly of ribosomes 1. Here, we report a quantitative label-free LC-MS/MS analysis to determine the nuclear proteome of Arabidopsis at 22ºC, HS (37ºC 4 and 24 hours), and a recovery phase. This analysis identified ten distinct groups of proteins based on their protein abundance in the nucleus before, during and after HS: the Early Group (EG), Late Group (LG), Transient Group (TG), Early Persistent Group (EPG), Late Persistent Group (LPG), Burst Group (BG), Early-Like Group (ELG), Late-Like Group (LLG), Transient-Like Group (TLG) and Continuous-Induced Group (CIG). Interestingly, the RNA Pol I subunit NRPA3 and other main nucleolar proteins – such as Nucleolin and Fibrillarin – were detected in the BG and CIG, suggesting that plants require increased nucleolar activity and likely ribosome assembly to restore protein synthesis after HS.
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Project description:Maintenance of the epigenome is of utmost importance to warrant appropriate cellular functioning Here we analyzed PRC1/2 function, in response to cellular stress. We observe that heat shock (HS) leads to nucleolar accumulation of CBX proteins, and are strongly immobilized in a 1,6-hexanediol insensitive manner, suggestive of a HS-induced liquid-to-solid phase transition of the nucleolus. CBX protein recovery from the nucleolus is dependent on heat shock protein (HSP) activity. LC-MS/MS analyses of nucleoli shows HS-induced nucleolar accumulation of PRC1/2 subunits, various chromatin regulators, HSPs, and the 26S proteasome. Finally, we find that HS leads to a strong reduction of PRC1/2 chromatin binding at target genes and loss of H2AK119ub and H3K27me3. Our findings demonstrate that thermal stress results in a rapid accumulation of epigenetic regulators in the nucleolus, and a concomitant loss of PRC1/2 complex chromatin binding and associated epimarks, underlining that environmental stress directly alters the epigenome.
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Project description:NF-YC10 is a subunit of the NF-Y transcription factor complex in Arabidopsis thaliana. We identified NF-YC10 as an interactor of the transcription factor DREB2A, which binds to DRE and activate expression of target genes under heat and/or dehydration stress conditions. Transgenic Arabidopsis plants that overexpress NF-YC10 showed stronger expression of DREB2A target genes under heat stress and were more tolerant to heat stress tolerance than the vector control plants. We conducted a transcript profiling by microarray aiming to identify target genes of NF-YC10 under heat stress
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