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Differences in RNA polymerase II complexes and their interactions with surrounding chromatin on human and cytomegalovirus genomes

ABSTRACT: Differences in RNA polymerase II complexes and their interactions with surrounding chromatin on human and cytomegalovirus genomes

PROVIDER: PRJNA770683 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16288500_1.fastq.gz	Fastqsanger.gz
	SRR16288500_2.fastq.gz	Fastqsanger.gz
	SRR16288501_1.fastq.gz	Fastqsanger.gz
	SRR16288501_2.fastq.gz	Fastqsanger.gz
	SRR16288502_1.fastq.gz	Fastqsanger.gz

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Differences in RNA polymerase II complexes and their interactions with surrounding chromatin on human and cytomegalovirus genomes [Pro-seq]

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Project description:To investigate the chromatin structure surrounding human RNA polymerase II (Pol II) transcription on host and viral genomes, chromatin immunoprecipitation (ChIP) was performed following digestion of HCMV infected primary human foreskin fibroblasts (HFFs) with DNA Fragmentation Factor (DFF). DFF is a human endonuclease responsible for cleaving between nucleosomes during apoptosis. We found that utilizing DFF as the front end for ChIP-Seq in place of sonication or Micrococcal nuclease (MNase) offered new insights into the connections between active transcription and the local chromatin on the host and that these connections were completely absent on the HCMV genome.

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Project description:Interactions of the RNA polymerase II (Pol II) preinitiation complex (PIC) and paused early elongation complexes with the first downstream (+1) nucleosome are thought to be functionally important. However, current methods are limited for investigating these relationships, both for cellular chromatin and the human cytomegalovirus (HCMV) genome. Digestion with human DNA fragmentation factor (DFF) before immunoprecipitation (DFF-ChIP) precisely revealed both similarities and major differences in PICs driven by TBP on the host genome in comparison with PICs driven by TBP or the viral-specific, late initiation factor UL87 on the viral genome. Host PICs and paused Pol II complexes are frequently found in contact with the +1 nucleosome and paused Pol II can also be found in a complex involved in the initial invasion of the +1 nucleosome. In contrast, viral transcription complexes have very limited nucleosomal interactions, reflecting a relative lack of chromatinization of transcriptionally active regions of HCMV genomes.

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Differences in RNA polymerase II complexes and their interactions with surrounding chromatin on human and cytomegalovirus genomes

Project description:This SuperSeries is composed of the SubSeries listed below.

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