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Genome-wide changes in active RNAPII occupancy in the presence of mutated TFIIS (TFIISmut)

ABSTRACT: Genome-wide changes in active RNAPII occupancy in the presence of mutated TFIIS (TFIISmut)

PROVIDER: PRJNA527114 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8730280.fastq.gz	Fastqsanger.gz
	SRR8730281.fastq.gz	Fastqsanger.gz
	SRR8730282.fastq.gz	Fastqsanger.gz
	SRR8730283.fastq.gz	Fastqsanger.gz
	SRR8730274.fastq.gz	Fastqsanger.gz

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Genome-wide changes in active RNAPII occupancy in the presence of mutated TFIIS (TFIISmut)

Project description:The expresion of dominant negative version of Arabidopsis TFIIS (TFIISmut) in plants lacking funtional TFIIS leads to severe growth defects. The genome-wide analysis of changes in active RNAPII occupancy upon TFIISmut expression was determined using ChIP-seq in order to better understand the biological importance of TFIIS. 10DAS seedlings of transgenic lines carring inducible GFP-TFIIS or GFP-TFIISmut transgenes were studied by ChIP-seq following plants exposure to 24h b-estradiol or EtOH (mock) induction.

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Transcriptomic changes in Arabidopsis thaliana upon the expression of mutated TFIIS

Project description:The expresion of dominant negative version of Arabidopsis TFIIS (TFIISmut) in plants lacking funtional TFIIS leads to severe growth defects. The genome-wide analysis of transcriptomic changes upon TFIISmut expression was performed by RNA-seq to better understand the biological importance of TFIIS. 10DAS seedlings of transgenic lines carring inducible GFP-TFIIS or GFP-TFIISmut transgenes were studied by RNA-seq following plants exposure to 24h b-estradiol or EtOH (mock) induction.

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Project description:Transcriptomic changes in Arabidopsis thaliana upon the expression of mutated TFIIS

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RNAPII ChIP-Sequencing of A.thaliana tfiis mutant lines under heat stress conditions

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RNAPII CTD dataset

Project description:The C-terminal domain (CTD) of RNA polymerase II (RNAPII) is composed of heptapeptide repeats, which play a key regulatory role in gene expression. Using genetic interaction and mRNA expression analysis we found that truncating the CTD resulted in distinct changes to cellular function. Truncating the CTD altered RNAPII occupancy to not only cause decreases, but also increases in mRNA levels. The latter were largely mediated by promoter elements, and in part were linked to the transcription factor Rpn4. The mediator subunit Cdk8 was enriched at promoters of these genes, and its removal not only restored normal mRNA and RNAPII occupancy levels, but also reduced the abnormally high cellular amounts of Rpn4. This suggested a positive role of Cdk8 in relationship to RNAPII, which contrasted with the negative role at the activated INO1 gene. Here, loss of CDK8 suppressed the reduced mRNA expression and RNAPII occupancy levels of CTD truncation mutants.

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Elongation Factor TFIIS Prevents Transcription Stress and R-Loop Accumulation to Maintain Genome Stability [ RNA-seq]

Project description:Connections between RNA polymerase II (RNAPII) transcription stress, R-loops, and genome instability have been established however, the underlying mechanisms remain poorly understood. Here we used a mutant version of elongation factor TFIIS (TFIISmut) to specifically induce increased levels of RNAPII pausing, arrest, and/or backtracking in human cells. TFIISmut expression results in slower elongation rates, relative depletion of polymerases from the end of genes, and increased levels of stopped RNAPII. It affects mRNA splicing and termination as well. Remarkably, however, TFIISmut expression also dramatically increases R-loops, which may form at the anterior end of backtracked RNAPII and trigger genome instability, including DNA strand breaks. These results shed new light on the relationship between transcription stress and R-loops, and suggest that different classes of R-loops exist, potentially with distinct consequences for genome instability.

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