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Genome-scale reconstruction of the sigma factor network in E. coli

ABSTRACT: We report the application of single-molecule-based sequencing technology for high-throughput profiling of transcription start sites for Escherichia coli under different conditions. By obtaining sequence from 5' RACE (rapid amplification of cDNA ends) followed by deep sequencing, we generated genome-wide TSS (transcription start site) maps for E. coli. This TSS-map was integrated with ChIP-chip data generated for 6 sigma factors in E. coli, resulting in reconstruction of sigma factor network in E. coli.

ORGANISM(S): Escherichia coli str. K-12 substr. MG1655

PROVIDER: GSE46737 | GEO | 2014/02/05

SECONDARY ACCESSION(S): PRJNA202224

REPOSITORIES: GEO

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Genome-scale reconstruction of the sigma factor network in E. coli

Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of transcription start sites for Escherichia coli under different conditions. By obtaining sequence from 5' RACE (rapid amplification of cDNA ends) followed by deep sequencing, we generated genome-wide TSS (transcription start site) maps for E. coli. This TSS-map was integrated with ChIP-chip data generated for 6 sigma factors in E. coli, resulting in reconstruction of sigma factor network in E. coli. Examination of transcription start sites by biological duplicates from E. coli for 3 different conditions

2014-02-05 | E-GEOD-46737 | biostudies-arrayexpress

Comparative analysis of regulatory elements between Escherichia coli and Klebsiella pneumoniae by genome-wide transcription start site profiling [TSS-Seq]

Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of transcription start sites for two enterobacteria: Escherichia coli and Klebsiella pneumoniae.By obtaining over fourteen billion bases of sequence from 5' RACE (rapid amplification of cDNA ends) followed by deep sequencing, we generated genome-wide TSS (transcription start site) maps for those two species. With experimentally derived TSS datasets, we examined usage of multiple TSSs, length of 5' UTR (untranslated region), SD (Shine-Dalgarno) sequence motif, promoter sequence motif, and dinucleotide preference near TSS site. In addition, we used the TSS datasets to identify sRNAs (small RNAs) in E. coli and K. pneumoniae. Based on these analysis, we compared regulatory elements including promoter, 5' UTR and sRNAs between two species, and investigated similarities and differences of upstream regulatory regions. Moreover, sRNAs were also compared and analyzed in terms of their sequences and target sequences, presenting possible working mechanisms of K. pneumoniae sRNAs by transferring prior knowledge from E. coli sRNAs. Examination of transcription start sites by biological duplicates from E. coli and K. pneumoniae

2012-07-30 | E-GEOD-35821 | biostudies-arrayexpress

Comparative analysis of regulatory elements between Escherichia coli and Klebsiella pneumoniae by genome-wide transcription start site profiling [TSS-Seq]

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Cappable-seq of Sinorhizobium meliloti ECF/anti-σ-free free strains overexpressing heterologous ecf genes compared to an empty vector control strain

Project description:Bacterial σ factors are dissociable subunits of the core RNA polymerase and important for promoter recognition and transcription initiation. Extracytoplasmic function (ECF) σ factors (σs) represent the most minimalistic and diverse group of alternative σs within the σ70 protein family. It has been shown that heterologous ECF σs hold great potential as context independent regulators in a variety of bacterial species and we implemented a core set of 12 heterologous ECF σs and their cognate promoters in Sinorhizobium meliloti. To analyze if heterologous ECF σs affect transcription from the host genome, we overexpressed the ecf02_2817 gene encoding σE from E. coli and the ecf11_0987 gene from Vibrio parahaemolyticus from ~20 copy number plasmid in RFF625c, a S. meliloti Rm1021 strain deleted for all native ECF/anti-σ genes (Lang et al. 2018, DOI: 10.1128/mSphereDirect.00454-18) . We employed Cappable-Seq to globally determine transcription start sites (TSS) in theses strains and compared it to the TSS profile of an empty vector control strain.

2023-01-26 | E-MTAB-12127 | biostudies-arrayexpress

The Global Landscape of Transcription Initiation During the Caulobacter Cell Cycle [mapping]

Project description:The Caulobacter cell cycle includes in an asymmetric cell division that is driven by a core regulatory circuit comprised of 4 transcription factors (DnaA, GcrA, CtrA, and SciP) and a DNA methyltransferase (CcrM). Using a modified global 5M-bM-^@M-^Y RACE protocol we mapped 2,726 transcriptional start sites (TSS) in the 4mb Caulobacter genome and identified 586 cell cycle-regulated TSS. The core cell cycle circuit directly controls about 55% of cell cycle-regulated TSS by integrating multiple regulatory inputs within at least 322 promoters, providing a large number of transcription profiles from a small number of regulatory factors. Here, we identified previously unknown features of the core cell cycle circuit, including antisense TSS within dnaA and ctrA, plus newly identified TSS for ctrA and ccrM. Altogether, we identified 615 antisense TSS plus 241 genes that are transcribed from multiple TSS. The multiple TSS in the same promoter region often exhibit different cell cycle activation timing, These novel features of the global transcript profile add significant insight to the system architecture of the Caulobacter cell cycle regulatory circuit. Global 5' RACE was performed to map Transcription Start Sites in the Caulobacter NA1000 genome

2014-10-16 | E-GEOD-57365 | biostudies-arrayexpress

The Global Landscape of Transcription Initiation During the Caulobacter Cell Cycle [timecourse]

Project description:The Caulobacter cell cycle includes in an asymmetric cell division that is driven by a core regulatory circuit comprised of 4 transcription factors (DnaA, GcrA, CtrA, and SciP) and a DNA methyltransferase (CcrM). Using a modified global 5M-bM-^@M-^Y RACE protocol we mapped 2,726 transcriptional start sites (TSS) in the 4mb Caulobacter genome and identified 586 cell cycle-regulated TSS. The core cell cycle circuit directly controls about 55% of cell cycle-regulated TSS by integrating multiple regulatory inputs within at least 322 promoters, providing a large number of transcription profiles from a small number of regulatory factors. Here, we identified previously unknown features of the core cell cycle circuit, including antisense TSS within dnaA and ctrA, plus newly identified TSS for ctrA and ccrM. Altogether, we identified 615 antisense TSS plus 241 genes that are transcribed from multiple TSS. The multiple TSS in the same promoter region often exhibit different cell cycle activation timing, These novel features of the global transcript profile add significant insight to the system architecture of the Caulobacter cell cycle regulatory circuit. Global 5' RACE was performed to measure Transcription Start Site activity at time points of the Caulobacter NA1000 cell cycle

2014-10-16 | E-GEOD-57364 | biostudies-arrayexpress

Characterization of Lactobacillus Sigma Factors using a Temperature Sensitive E. coli Sigma70 Mutant

Project description:Housekeeping sigma factors in the Sigma70 family, as components of the RNA polymerase holoenzyme, are responsible for regulating transcription of genes related to vegetative growth. While these factors are well understood in model organisms such as Escherchia coli and Bacillus subtilis, little experimental work has focused on the sigma factors in members of the Lactobacillus genus such as Lactobacillus brevis and Lactobacillus plantarum. This study evaluates the ability of putative Sigma70 proteins from L. brevis (Sigma70-Lb) and L. plantarum (Sigma70-Lp) to complement a temperature sensitive mutation in E. coli 285c Sigma70. After finding that the heterologous sigma factors were capable of restoring the viability of E. coli 285c at 42 C through growth kinetics studies, the transcriptional responses of 285c to an extended heat shock in the presence of Sigma70-Lb and Sigma70-Lp were found to be similar to previous studies. These results indicate the Sigma70-Lb and Sigma70-Lp are capable of initiating transcription in a complex with the E. coli 285c RNA polymerase to a sufficient degree to restore viability at elevated temperatures without triggering unusual modifications to the native transcriptional program. These heterologous sigma factors may therefore be useful to improve biochemical knowledge of the sigma factor family or for use in transcriptional engineering.

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Identification and validation of antisense RNAs in Escherichia coli

Project description:In this study transcriptional start sites (TSS) of E. coli were determined for several growth conditions

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Escherichia coli Sigma 32 ChIP-chip

Project description:Identification of Sigma 32 binding sites across the Escherichia coli genome using ChIP-chip following heat shock.

2013-09-03 | E-MTAB-1849 | biostudies-arrayexpress

Time course analysis of genes regulated by E. coli sigma 32 (rpoH) after overexpression

Project description:DNA microarray analysis of genes regulated by the alternative sigma factor, sigma 32 (rpoH). Sigma 32-dependent genes were initially identified by comparing a wild type (wt) E. coli K-12 strain that has a low level of sigma 32, with a strain over-expressing sigma 32 (following induction of rpoH from an inducible promoter by IPTG). We monitored changes in gene expression in 4 separate time-courses. Because sigma 32 is negatively regulated, its activity decreases 10 min after overexpression. Consequently, to identify genes regulated by sigma 32 we analyzed the expression data for 10 min after overexpression using SAM (Statistical Analysis of Microarrays). We identified 105 genes organized in 66 separate transcription units. This study is detailed in Nonaka et al 2006 (submitted). Keywords: time course

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