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SRX7255737: GSM4200370: Rpo47.expon.chip-seq.r2; Saccharolobus solfataricus; ChIP-Seq
1 ILLUMINA (Illumina HiSeq 4000) run: 36.8M spots, 5.5G bases, 1.8Gb downloads

Submitted by: NCBI (GEO)
Study: Pervasive promoter-proximal regulation in archaea: ChIP-seq exponential growth
show Abstracthide Abstract
Recruitment of RNA polymerase and initiation factors to the promoter is the only known mechanisms for transcription activation and repression in archaea. Whether any of the subsequent steps towards productive transcription elongation is involved in regulation is not known. We characterised how the basal transcription machinery is distributed along genes in the archaeon Sulfolobus solfataricus. We discovered a distinct early elongation phase where RNA polymerases sequentially recruit the elongation factors Spt4/5 and Elf1 to form the transcription elongation complex (TEC) before the TEC escapes into productive transcription. TEC escape is rate-limiting for transcription output during exponential growth. Oxidative stress causes changes in TEC escape that correlate with changes in the transcriptome. Our results thus establish that TEC escape contributes to the basal promoter strength and facilitates transcription regulation. Impaired TEC escape coincides with the accumulation of initiation factors at the promoter and recruitment of termination factor aCPSF1 to the early TEC. This suggests two possible mechanisms for how TEC escape limits transcription, physically blocking upstream RNA polymerases during transcription initiation and premature termination of early TECs. Overall design: ChIP-seq with polyclonal antibodies
Sample: Rpo47.expon.chip-seq.r2
SAMN13446983 • SRS5753275 • All experiments • All runs
Library:
Instrument: Illumina HiSeq 4000
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: PAIRED
Construction protocol: Cells were lysed in lysis buffer (50 mM HEPES/NaOH pH 7.5, 140 mM NaCl, 1 mM EDTA, 0.1% Na-deoxycholate, 1% Triton X-100) and DNA was sheared by sonication. Lysates were cleared by centrifugation. Chromatin-immunoprecipitation was carried out with protein-A-purified polyclonal antibodies. Immunoprecipitated DNA was recovered by QIAquick PCR purification NEBnext Ultra II DNA library preparation
Experiment attributes:
GEO Accession: GSM4200370
Links:
Runs: 1 run, 36.8M spots, 5.5G bases, 1.8Gb
Run# of Spots# of BasesSizePublished
SRR1057430236,834,1645.5G1.8Gb2020-09-16

ID:
9527801

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