Curation Information

Publication
ChIP-seq and transcriptome analysis of the OmpR regulon of Salmonella enterica serovars Typhi and Typhimurium reveals accessory genes implicated in host colonization.;Perkins TT, Davies MR, Klemm EJ, Rowley G, Wileman T, James K, Keane T, Maskell D, Hinton JC, Dougan G, Kingsley RA;Molecular microbiology 2013 Feb; 87(3):526-38 [23190111]
TF
OmpR [P0AA16, view regulon]
Reported TF sp.
Salmonella enterica subsp. enterica serovar Typhi str. Ty2
Reported site sp.
Salmonella enterica subsp. enterica serovar Typhi str. Ty2
Created by
Elliot White
Curation notes
-

Experimental Process

Researchers used CHiP-seq to discover OmpR binding sites; binding sequences were determined from enriched sequences using the motif finding algorithm YMF. Regulation was assayed in a previous paper (19609351) with both a microarray and RNA-seq.

ChIP assay conditions
Normal cell conditions; ompR binding was not induced.
ChIP notes
Salmonella Typhi ompR::3×FLAG (strain TT53) and S. Typhi BRD948 were cultured in LB broth to OD600 = 0.6, lysed, incubated with 1% formaldehyde at 37°C for 20 min to cross-link DNA with protein then quenched with glycine (ph7) to a final concentration of 0.5 M. Cells were harvested and washed twice in TBS and lysed by osmotic shock. Genomic DNA was then sheered by sonication to an average size of 300 bp and immunoprecipitated using anti-3×FLAG monoclonal antibody (Sigma, F3165) as previously described (Pfeiffer et al., 2007) using the Protein G Immunoprecipitation kit (Sigma). Eluates were then treated with pronase (0.8 mg ml−1, Sigma) at 65°C overnight. The nucleotide sequence of genomic DNA fragments was determined by Illumina GAII paired-end sequencing with read length 36 bp and mapped to the S. Typhi Ty2 whole genome sequence (AE014613). Sequence data were mapped to the S. Typhi Ty2 genome using the same parameters as previously described (Perkins et al., 2009), without assigning the sequence reads to each strand. Plots were z-score-normalized, in order to indicate the number of standard deviations above or below the mean for each datum point, and the differences between the untagged S. Typhi Ty2 and ompR::3×FLAG-tagged associated DNA sequences determined. Plots were then read into the genome browser tool Artemis (Rutherford et al., 2000). The Peakfinder function was used to determine enrichment for OmpR::3×FLAG bound DNA sequences. The Peakfinder function (36 bp window and z-score cut-off score set to 3) identified 58 peaks. Due to the background noise of the mapped sequence data plots and low stringency of the Peakfinder conditions, identified peaks were then filtered manually. Sites of DNA enrichment present within a predicted or known CDS were ignored unless there were multiple similar sites nearby, reducing the total number of analysed peaks to 43. Enriched sequences were then input to the motif finding algorithm YMF with the length of motif set to eight nu

Transcription Factor Binding Sites


TAGGCCA
AGTTAGT
TGTTAGT
ATGTAAC
TGTAACA
TGTTACA
TAGGCCA
TTGTTAC
TGTTACA
GGCTAGA
ATGTAAC
TTGTTAC
TGTTACA
ATGTAAC
ATGTTAC
TGTTACA
ATGTTAC
AGTTAGT
ACGGCCT
TTGTAAC
TGTAACA
TGTAACA
TTGTAAC
TGTAACA
AACTAAC
ACGGCCT
TAGTCTA
TGTTACA
TGTTAGT
CGCTAGA
ATGTAAC
TGTAACA
TGTAACA
TAGGCCA
AGTTAGT
TGTTAGT
ATGTAAC
TGTAACA
TGTTACA
TAGGCCA
TTGTTAC
TGTTACA
GGCTAGA
ATGTAAC
TTGTTAC
TGTTACA
ATGTAAC
ATGTTAC
TGTTACA
ATGTTAC
AGTTAGT
ACGGCCT
TTGTAAC
TGTAACA
TGTAACA
TTGTAAC
TGTAACA
AACTAAC
ACGGCCT
TAGTCTA
TGTTACA
TGTTAGT
CGCTAGA
ATGTAAC
TGTAACA
TGTAACA

Quantitative data format: Peak intensity; called by PeakFinder

TAGGCCA 7.31772
AGTTAGT 6.4539347
TGTTAGT 6.4539347
ATGTAAC 6.4539347
TGTAACA 6.4539347
TGTTACA 6.4539347
TAGGCCA 7.31772
TTGTTAC 8.895602
TGTTACA 6.4539347
GGCTAGA 7.7002826
ATGTAAC 6.4539347
TTGTTAC 8.895602
TGTTACA 6.4539347
ATGTAAC 6.4539347
ATGTTAC 4.129863
TGTTACA 6.4539347
ATGTTAC 4.129863
AGTTAGT 6.4539347
ACGGCCT 6.789148
TTGTAAC 7.7002826
TGTAACA 6.4539347
TGTAACA 6.4539347
TTGTAAC 7.7002826
TGTAACA 6.4539347
AACTAAC 6.4539347
ACGGCCT 6.789148
TAGTCTA 8.895602
TGTTACA 6.4539347
TGTTAGT 6.4539347
CGCTAGA 16.233952
ATGTAAC 6.4539347
TGTAACA 6.4539347
TGTAACA 6.4539347

Gene Regulation

Regulated genes for each binding site are displayed below. Gene regulation diagrams show binding sites, positively-regulated genes, negatively-regulated genes, both positively and negatively regulated genes, genes with unspecified type of regulation. For each indvidual site, experimental techniques used to determine the site are also given.

Site sequence Regulated genes Gene diagram Experimental techniques TF function TF type
TAGGCCA
... ... t0528 ackA pta t0529 t0530 t0531 t0532 t0533
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
AGTTAGT
... ... galS yeiB folE mglB mglA mglC yeiA t0669
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTTAGT
... ... galS yeiB folE mglB mglA mglC yeiA t0669
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
ATGTAAC
... ... ompS t0884 yedJ dcm vsr yedA
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTAACA
... ... ompS t0884 yedJ dcm vsr yedA
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTTACA
... ... tppB gst pdxY tyrS pdxH ydhA anmK nth ydgQ t1323 rnfD t1325 t1326 t1327 t1328 t1329
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TAGGCCA
... ... tppB gst pdxY tyrS pdxH ydhA anmK nth ydgQ t1323 rnfD t1325 t1326 t1327 t1328 t1329
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TTGTTAC
... ... csgB csgA csgC t1774 t1773 t1772 mdoC csgD csgE csgF csgG t1782 t1783 t1784 t1785
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTTACA
... ... csgB csgA csgC t1774 t1773 t1772 mdoC csgD csgE csgF csgG t1782 t1783 t1784 t1785
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
GGCTAGA
... ... csgB csgA csgC t1774 t1773 t1772 mdoC csgD csgE csgF csgG t1782 t1783 t1784 t1785
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
ATGTAAC
... ... ompX ybiF
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TTGTTAC
... ... sdhC sdhD sdhA sdhB t2145 gltA t2147 nei t2149 t2150 t2151
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTTACA
... ... sdhC sdhD sdhA sdhB t2145 gltA t2147 nei t2149 t2150 t2151
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
ATGTAAC
... ... t2145 sdhC sdhD sdhA sdhB gltA t2147 nei t2149 t2150 t2151
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
ATGTTAC
... ... metK t05990 galP t3004 endA t3006 gshB
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTTACA
... ... metK t05990 galP t3004 endA t3006 gshB
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
ATGTTAC
... ... t3143 ygjQ t3144
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
AGTTAGT
... ... t3884 t3883 t3882 t3881 t3880 yhjW dppA
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
ACGGCCT
... ... t3884 t3883 t3882 t3881 t3880 yhjW dppA
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TTGTAAC
... ... t3884 t3883 t3882 t3881 t3880 yhjW dppA
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTAACA
... ... t3884 t3883 t3882 t3881 t3880 yhjW dppA
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTAACA
... ... yhjE t3907 yhjH yhjD yhjC
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TTGTAAC
... ... greB t4002 ompR envZ pckA
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTAACA
... ... greB t4002 ompR envZ pckA
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
AACTAAC
... ... tviA t4354 t4355 t4356
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
ACGGCCT
... ... tviA t4354 t4355 t4356
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TAGTCTA
... ... tviA t4354 t4355 t4356
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTTACA
... ... tviA t4354 t4355 t4356
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTTAGT
... ... tviA t4354 t4355 t4356
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
CGCTAGA
... ... tviA t4354 t4355 t4356
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
ATGTAAC
... ... t4356 t4355 t4354 t4357
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTAACA
... ... t4356 t4355 t4354 t4357
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified
TGTAACA
... ... t4356 t4355 t4354 t4357
Experimental technique details ChIP-Seq (ECO:0006009) - Experimental technique details Motif-discovery (ECO:0005558) - not specified not specified