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Continuous transcription initiation guarantees robust repair of all transcribed genes and regulatory regions

ABSTRACT: Continuous transcription initiation guarantees robust repair of all transcribed genes and regulatory regions

PROVIDER: PRJNA515483 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8448030.fastq.gz	Fastqsanger.gz
	SRR8448040.fastq.gz	Fastqsanger.gz
	SRR8448031.fastq.gz	Fastqsanger.gz
	SRR8448041.fastq.gz	Fastqsanger.gz
	SRR8448032.fastq.gz	Fastqsanger.gz

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Continuous transcription initiation guarantees robust repair of all transcribed genes and regulatory regions

Project description:Inhibition of transcription caused by DNA damage-impaired RNA polymerase II (Pol II) elongation conceals a local increase in de novo transcription, slowly progressing from Transcription Start Sites (TSSs) to gene ends. Although associated with accelerated repair of Pol II-encountered lesions and limited mutagenesis, it is still unclear how this mechanism is maintained during genotoxic stress-recovery. Here we uncover a widespread gain in chromatin accessibility and preservation of the active H3K27ac mark after UV-irradiation. The concomitant increase in Pol II escape from promoter-proximal pause (PPP) sites of most active genes, PROMPTs and enhancer RNAs favors unrestrained initiation, as evidenced by the synthesis of nascent RNAs including start RNAs. Accordingly, drug-inhibition of PPP-release replenishes levels of pre-initiating Pol II at TSSs after UV. Our data show that such continuous engagement of Pol II molecules ensures maximal transcription-driven repair throughout expressed genes and regulatory loci. Importantly, revealing this unanticipated regulatory layer of UV-response provides physiological relevant traction to the emerging concept that Pol II initiation rate is determined by pause-release dynamics.

2020-02-14 | GSE125181 | GEO

Continuous transcription initiation guarantees robust repair of all transcribed genes and regulatory regions.

| S-EPMC7021815 | biostudies-literature

Genome-wide depletion of replication initiation events in highly transcribed regions.

Project description:This report investigates the mechanisms by which mammalian cells coordinate DNA replication with transcription and chromatin assembly. In yeast, DNA replication initiates within nucleosome-free regions, but studies in mammalian cells have not revealed a similar relationship. Here, we have used genome-wide massively parallel sequencing to map replication initiation events, thereby creating a database of all replication initiation sites within nonrepetitive DNA in two human cell lines. Mining this database revealed that genomic regions transcribed at moderate levels were generally associated with high replication initiation frequency. In genomic regions with high rates of transcription, very few replication initiation events were detected. High-resolution mapping of replication initiation sites showed that replication initiation events were absent from transcription start sites but were highly enriched in adjacent, downstream sequences. Methylation of CpG sequences strongly affected the location of replication initiation events, whereas histone modifications had minimal effects. These observations suggest that high levels of transcription interfere with formation of pre-replication protein complexes. Data presented here identify replication initiation sites throughout the genome, providing a foundation for further analyses of DNA-replication dynamics and cell-cycle progression.

| S-EPMC3205567 | biostudies-literature

Ssu72 is essential to suppress initiation of cryptic transcription in the 3' end of the RNA polymerase II transcribed regions.

Project description:Ssu72 is essential to suppress initiation of cryptic transcription in the 3' end of the RNA polymerase II transcribed regions.

| PRJNA604184 | ENA

Transcription shapes DNA replication initiation to preserve genome integrity

Project description:We develop a high-throughput nucleoside analog incorporation sequencing assay and identify thousands of early replication initiation zones (ERIZs) in both mouse and human cells. The identified ERIZs fall in open chromatin compartments while are mutually exclusive with transcription elongation and occupy mainly non-transcribed regions adjacent to transcribed regions. Furthermore, we reveal that RNA polymerase II actively redistributes the chromatin-encircled mini-chromosome maintenance (MCM) complex but not the origin-recognition complex (ORC) to actively restrict early DNA replication initiation outside of transcribed regions. The coupling of RNA polymerase II and MCM is further validated by detected MCM accumulation and DNA replication initiation after RNA polymerase II stalling via anchoring nuclease-dead Cas9 at the transcribed genes. Importantly, we also find that the orchestration of DNA replication initiation by transcription can efficiently prevent gross DNA damage.

2021-05-20 | GSE174680 | GEO

Transcribed Ultraconserved Regions in Cancer.

Project description:Transcribed ultraconserved regions are putative lncRNA molecules that are transcribed from DNA that is 100% conserved in human, mouse, and rat genomes. This is notable, as lncRNAs are typically poorly conserved. TUCRs remain very understudied in many diseases, including cancer. In this review, we summarize the current literature on TUCRs in cancer with respect to expression deregulation, functional roles, mechanisms of action, and clinical perspectives.

| S-EPMC9139194 | biostudies-literature

Screening thousands of transcribed coding and non-coding regions reveals sequence determinants of RNA polymerase II elongation potential.

Project description:Organismal growth and development rely on RNA Polymerase II (RNAPII) synthesizing the appropriate repertoire of messenger RNAs (mRNAs) from protein-coding genes. Productive elongation of full-length transcripts is essential for mRNA function, however what determines whether an engaged RNAPII molecule will terminate prematurely or transcribe processively remains poorly understood. Notably, despite a common process for transcription initiation across RNAPII-synthesized RNAs1, RNAPII is highly susceptible to termination when transcribing non-coding RNAs such as upstream antisense RNAs (uaRNAs) and enhancers RNAs (eRNAs)2, suggesting that differences arise during RNAPII elongation. To investigate the impact of transcribed sequence on elongation potential, we developed a method to screen the effects of thousands of INtegrated Sequences on Expression of RNA and Translation using high-throughput sequencing (INSERT-seq). We found that higher AT content in uaRNAs and eRNAs, rather than specific sequence motifs, underlies the propensity for RNAPII termination on these transcripts. Further, we demonstrate that 5’ splice sites exert both splicing-dependent and autonomous, splicing-independent stimulation of processive transcription, even in the absence of polyadenylation signals. Together, our results reveal a potent role for transcribed sequence in dictating gene output at mRNA and non-coding RNA loci, and demonstrate the power of INSERT-seq towards illuminating these contributions.

2022-04-06 | GSE178230 | GEO

Transcription profiling of human circulating blood reticulocytes from adult and cord blood to provide a robust description of genes transcribed at the final stages of erythroblast maturation

Project description:RNA from circulating blood reticulocytes was utilized to provide a robust description of genes transcribed at the final stages of erythroblast maturation. After depletion of leukocytes and platelets, Affymetrix HG-U133 arrays were hybridized with probe from total RNA isolated from blood sampled from 14 umbilical cords and 14 healthy adult humans. Experiment Overall Design: Peripheral blood from 14 normal adult and 14 human umbilical cord were purified to obtain packed red blood cells and their purity was assessed by CELL-DYN4000. The contaminated WBC count was less than 1 out of 1 million cells. Total RNA from purified reticulocytes included in the purified red blood cells was extracted, labeled, and hybridized onto Affymetrix HG-U133 A and B arrays.

2007-11-01 | E-GEOD-6236 | biostudies-arrayexpress

Continuous thermophilic composting

Project description:Continuous thermophilic composting Raw sequence reads

| PRJNA833274 | ENA

Crosslinking mass spectrometry of yeast Pol I pre-initiation complex

Project description:In eukaryotic cells, ribosomal RNA is transcribed by RNA polymerase I (Pol I). To initiate transcription, Pol I requires the assembly of a multi-subunit pre-initiation complex (PIC). In yeast, the minimal PIC includes Pol I, the transcription factor Rrn3 and Core Factor (CF) composed of subunits Rrn6, Rrn7 and Rrn11. We have determined the cryo-EM structure of a 18-subunit yeast Pol I complex together with CF, Rrn3 and transcription scaffold. To aid in modeling the structure based on the cryo-EM map, we performed crosslinking mass spectrometry.

2017-07-31 | PXD006510 | Pride

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