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SRX360044: GSM1239612: batch5_chrom2_LoVo_THAP4_Mouse_FailedQC; Homo sapiens; ChIP-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 26.8M spots, 856.8M bases, 496.3Mb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Transcription factor binding in human cells occurs in dense clusters formed around cohesin anchor sites [Validation Set]
show Abstracthide Abstract
During cell division, transcription factors (TFs) are removed from chromatin twice, during DNA synthesis, and during condensation of chromosomes. How TFs can efficiently find their sites following these stages has been unclear. Here, we have analyzed the binding pattern of expressed TFs in human colorectal cancer cells. We find that binding of TFs is highly clustered, and that the clusters are enriched in binding motifs for several major TF classes. Strikingly, almost all clusters are formed around cohesin, and loss of cohesin decreases both DNA accessibility and binding of TFs to clusters. We show that cohesin remains bound in S phase, holding the nascent sister chromatids together at the TF cluster sites. Furthermore, cohesin remains bound to the cluster sites when TFs are evicted in early M-phase. These results suggest that cohesin binding functions as a cellular memory that promotes re- stablishment of TF clusters after DNA replication and chromatin condensation. Overall design: Examination of TF binding by ChIP-seq in a CRC cell-line.
Sample: batch5_chrom2_LoVo_THAP4_Mouse_FailedQC
SAMN02363725 • SRS487798 • All experiments • All runs
Organism: Homo sapiens
Library:
Instrument: Illumina HiSeq 2000
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: Cells were cross-linked by 1% formaldehyde, and DNA was sonicated to 200- 400 bp fragments. Antibodies (5 µg) were added and collected using protein G Sepharose (GE). Cross-links were reversed and proteins digested by incubation with proteinase K at 65 °C overnight. DNA was then purified using Qiagen PCR purification kit. Libraries were constructed according to Illuminas standard protocol. The concentration of the library was determined by Nanodrop spectrophotometer (Thermo Fisher Scientific Inc.) and 10 pmol of the DNA was applied for one flow-cell lane. Sequencing was performed using one lane of Illumina GAIIx, or alternatively, samples were indexed with DNA barcodes and four to six different samples were multiplexed in one library and sequenced using Illumina HiSeq 2000 (single 36 bp reads).
Experiment attributes:
GEO Accession: GSM1239612
Links:
External link:
Runs: 1 run, 26.8M spots, 856.8M bases, 496.3Mb
Run# of Spots# of BasesSizePublished
SRR100257926,775,147856.8M496.3Mb2013-09-26

ID:
509247

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