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SRX18804820: GSM6876593: ZM4, RM, rep1; Zymomonas mobilis; Hi-C
1 ILLUMINA (HiSeq X Ten) run: 269.5M spots, 80.9G bases, 28.4Gb downloads

External Id: GSM6876593_r1
Submitted by: Biogas Institute of Ministry of Agriculture and Rural Affairs
Study: Transcription factor shapes chromosomal conformation and regulates gene expression in bacterial adaptation [Hi-C]
show Abstracthide Abstract
Genomic mutations allow bacteria to adapt rapidly to adverse stress environments. The three-dimensional conformation of the genome also may plays an important role in transcriptional regulation and environmental adaptation. Here, using chromosome conformation capture, we investigate the high-order architecture of the Zymomonas mobilis chromosome in response to genomic mutant and ambient stimuli (acetic acid and furfural, derived from lignocellulosic hydrolysate). We find that genomic mutation only influences the local chromosome contacts, whereas stress of acetic acid and furfural restrict the long-range contacts and change the chromosome organization at domain scales significantly. Further deciphering the domain feature unveils the important transcription factors, Ferric uptake regulation (Fur) proteins, which act as nucleoid-associated proteins to promote long-range (> 200 kb) chromosomal communications and regulate the expression of genes involved in stress response. Our work suggests that ubiquitous transcription factors in prokaryotes mediate chromosome organization and regulate stress-resistance genes in bacterial adaptation. Hi-C analysis revealed the three-dimensional conformation of Zymomonas mobilis. Overall design: The chromosomal conformation analysis of wild-type strain (ZM4-RM), genomic mutation strain (ZM532-RM), genomic mutation strain under environmental stress (ZM532-AF), and strains with the deletion of Fur proteins (zur and zurfur).
Sample: ZM4, RM, rep1
SAMN32348344 • SRS16238056 • All experiments • All runs
Library:
Name: GSM6876593
Instrument: HiSeq X Ten
Strategy: Hi-C
Source: GENOMIC
Selection: other
Layout: PAIRED
Construction protocol: Cells were then crosslinked with 3% fresh formaldehyde for 30 min at room temperature, followed by 30 min at 4 °C. Formaldehyde was finally quenched with 0.375 M glycine for 20 minutes at 4 °C. Fixed cells were collected, frozen in liquid nitrogen, and stored at -80 °C until use. 1 × 10^9 cells were suspended in 100 μl TE with 2 μl of Ready-Lyse lysozyme (Epicentre), and incubated at room temperature for 20 min before being treated with 0.5% SDS treatment for 10 min at 65 °C. SDS was quenched by adding 50 μl 10% Triton X-100 and incubating it for 15 min at 37 °C. The chromosomal DNA in the cell lysate was then digested for 1 h at 37 °C by adding 50 μl 10x NEB buffer 2.1(NEB), 300μl water, and 100U of Sau3AI. Biotin-14-dCTP (TriLINK) was used to label the restriction fragment ends with biotinylated cytosine nucleotides. Labeled DNA was diluted in a 10.0 ml ligation mix containing 100 Weiss units of T4 DNA ligase (Thermo) and ligated overnight at 16 °C. The crosslinking was reversed overnight at 65°C with 200 μg ml-1 proteinase K (Thermo), and DNA was purified using QIAamp DNA Mini Kit (Qiagen) according to the manufacturer's instructions. Purified DNA was sheared to a length of ~400 bp. Biotinylated DNA fragments were pulled down by Dynabeads MyOne Streptavidin C1 beads (Thermo fisher) and prepared for Illumina sequencing by NEBNext Ultra II DNA Library Prep Kit (NEB) as per manufacturer's instructions. The DNA fragments between 400 and 600 bp were purified and paired-end sequenced on the Illumina HiSeq X Ten platform with 150PE mode.
Runs: 1 run, 269.5M spots, 80.9G bases, 28.4Gb
Run# of Spots# of BasesSizePublished
SRR22845754269,542,07880.9G28.4Gb2024-05-16

ID:
25894223

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