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SRX885666: GSM1616440: rpoA_empty2; Pseudomonas aeruginosa; ChIP-Seq
1 ILLUMINA (Illumina HiSeq 2500) run: 14.1M spots, 703.6M bases, 457Mb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Chromatin Immunoprecipitation of HA-tagged PA14_69770 and RpoA in Pseudomonas aeruginosa UCBPP-PA14
show Abstracthide Abstract
Extremely slow growth imposed by energy limitation is a ubiquitous but poorly understood physiological state for microbes. We used oxygen limitation to impose this state on Pseudomonas aeruginosa and measured newly synthesized proteins using a time-selective proteome labeling method (BONCAT) to identify relevant regulators and metabolic pathways. We further characterized one upregulated protein that has no homology to any known protein domains. This small, acidic protein is post-transcriptionally regulated and physically interacts with RNA polymerase, binding near the secondary channel during transcription elongation, and leading to widespread effects on gene expression. For some genes, the impacts on transcript and protein levels are different, suggesting possible modulation of translation as well. These effects have phenotypic consequences, as deletion of the gene affects biofilm formation, secondary metabolite production, and fitness in fluctuating conditions. Based on these phenotypes, we have designated the protein SutA (survival under transitions). Overall design: Profiles of genome association of HA-tagged PA14_69770 or RpoA in cells lacking or overexpressing PA14_69770, grown to late stationary phase in minimal medium containing pyruate as the carbon source, in triplicate.
Sample: rpoA_empty2
SAMN03365902 • SRS852572 • All experiments • All runs
Library:
Instrument: Illumina HiSeq 2500
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: Pellets were lysed by treatment with 1 mg/ml lysozyme and sonicated with a microtip sonicator for 4 minutes, alternating 30 seconds on and 30 seconds off to shear DNA. Immunoprecipitation with an anti-HA antibody pre-conjugated to agarose beads or with an anti-RpoA antibody followed by protein A/G-conjugated agarose beads was carried out as descibed by Gilbert et al (Molecular microbiology 73, 1072-1085). Extracted, RNAse-treated DNA was purified using QIAquick columns and used for library construction. Libraries for ChIP Seq were constructed from the fragmented genomic DNA samples using the NEBNext ChIP Library Prep Master Mix Set for Illumina, according to the manufacturer's instructions.
Experiment attributes:
GEO Accession: GSM1616440
Links:
External link:
Runs: 1 run, 14.1M spots, 703.6M bases, 457Mb
Run# of Spots# of BasesSizePublished
SRR181321014,071,625703.6M457Mb2015-06-01

ID:
1272525

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