Limits...
Pilicide ec240 disrupts virulence circuits in uropathogenic Escherichia coli.

Greene SE, Pinkner JS, Chorell E, Dodson KW, Shaffer CL, Conover MS, Livny J, Hadjifrangiskou M, Almqvist F, Hultgren SJ - MBio (2014)

Bottom Line: Furthermore, the effect of ec240 on motility was abolished in the absence of the SfaB, PapB, SfaX, and PapX regulators.In contrast to the effects of ec240, deletion of the type 1 pilus operon led to increased S and P piliation and motility.Thus, ec240 dysregulated several uropathogenic Escherichia coli (UPEC) virulence factors through different mechanisms and independent of its effects on type 1 pilus biogenesis and may have potential as an antivirulence compound.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Microbiology and Microbial Pathogenesis and Center for Women's Infectious Disease Research, Washington University School of Medicine in St. Louis, St. Louis, Missouri, USA.

Show MeSH

Related in: MedlinePlus

Global transcriptional and proteomic responses to ec240. (A) Genes whose expression is dysregulated by growth in ec240 rather than DMSO, as identified by RNA-Seq analysis. Genes were classified by using KEGG (http://www.genome.jp/kegg/) and EcoCyc (http://www.ecocyc.org). The genes in each functional category are shown as percentages of the total genes dysregulated. (B) MA plots of UTI89 gene abundance following treatment with ec240. Each dot represents an annotated UTI89 gene with the log2 of its relative abundance in ec240 versus that in DMSO (M) plotted against the average log2 of its abundance under both conditions (A). M and A values are based on data from three biological replicates under each growth condition. The dotted lines indicate 3-fold changes. (C) Fold changes in CUP gene transcripts altered in UTI89 grown with ec240 rather than the DMSO vehicle control, as determined by RNA-Seq. Genes are shown in their order within the operon. (D) Proteins dysregulated after growth in ec240 rather than DMSO, as determined by iTRAQ analysis. Proteins were classified by using KEGG (http://www.genome.jp/kegg/) and EcoCyc (http://www.ecocyc.org). The proteins in each functional category are shown as percentages of the total proteins dysregulated and include proteins with increased and decreased abundances. (E) Fold changes in CUP proteins altered in UTI89 grown with ec240 rather than the DMSO vehicle control, as determined by iTRAQ analysis. (F) Dysregulation by ec240 of siderophore-related and iron transport genes by both RNA-Seq and iTRAQ analysis. Shown are fold changes in transcript or protein levels in UTI89 grown in ec240 rather than DMSO. Genes are shown in their order within the operon.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4217179&req=5

fig2: Global transcriptional and proteomic responses to ec240. (A) Genes whose expression is dysregulated by growth in ec240 rather than DMSO, as identified by RNA-Seq analysis. Genes were classified by using KEGG (http://www.genome.jp/kegg/) and EcoCyc (http://www.ecocyc.org). The genes in each functional category are shown as percentages of the total genes dysregulated. (B) MA plots of UTI89 gene abundance following treatment with ec240. Each dot represents an annotated UTI89 gene with the log2 of its relative abundance in ec240 versus that in DMSO (M) plotted against the average log2 of its abundance under both conditions (A). M and A values are based on data from three biological replicates under each growth condition. The dotted lines indicate 3-fold changes. (C) Fold changes in CUP gene transcripts altered in UTI89 grown with ec240 rather than the DMSO vehicle control, as determined by RNA-Seq. Genes are shown in their order within the operon. (D) Proteins dysregulated after growth in ec240 rather than DMSO, as determined by iTRAQ analysis. Proteins were classified by using KEGG (http://www.genome.jp/kegg/) and EcoCyc (http://www.ecocyc.org). The proteins in each functional category are shown as percentages of the total proteins dysregulated and include proteins with increased and decreased abundances. (E) Fold changes in CUP proteins altered in UTI89 grown with ec240 rather than the DMSO vehicle control, as determined by iTRAQ analysis. (F) Dysregulation by ec240 of siderophore-related and iron transport genes by both RNA-Seq and iTRAQ analysis. Shown are fold changes in transcript or protein levels in UTI89 grown in ec240 rather than DMSO. Genes are shown in their order within the operon.

Mentions: We assessed the transcriptional profile of UTI89 grown with 250 µM ec240 or a dimethyl sulfoxide (DMSO) vehicle control to investigate the global effects of ec240 on UPEC biology. We utilized a relatively high concentration of ec240 in order to detail all possible “off-target” effects. RNA-Seq was conducted on triplicate biological samples grown under type 1 pilus-inducing conditions. We identified 52 gene transcripts that were altered ≥3-fold when UTI89 was grown in ec240 rather than the DMSO vehicle control. Of these 52 genes, 15 had decreased transcript levels and 37 had increased transcript levels. The transcript levels of CUP pilus genes were affected by the pilicide ec240, as were the transcript levels of genes with a variety of different functions, including motility, siderophore synthesis and transport, metabolism, translation, gene regulation, and stress response (Fig. 2A). There were also some hypothetical genes with dysregulated transcript levels (Fig. 2A). As shown in the MA plot (a plot of the log2 of the ratio of abundances of each transcript between the two conditions [M] plotted against the average log2 of abundance of that transcript in both conditions [A]), the pap and sfa genes encoding P and S pili had the most upregulated transcript levels, while the fim and flg genes encoding type 1 pili and flagella had the most downregulated transcript levels (Fig. 2B), with fim transcripts downregulated 8- to 17-fold, pap transcripts upregulated 4- to 49-fold, and sfa transcripts upregulated 4- to 25-fold (Fig. 2C). Furthermore, CUP operon genes cumulatively make up 46% of the genes dysregulated by >3-fold but 76% of the genes dysregulated by >10-fold after growth in ec240. This indicates that under type 1 pilus-inducing conditions, the major effect of ec240 is the dysregulation of CUP pili.


Pilicide ec240 disrupts virulence circuits in uropathogenic Escherichia coli.

Greene SE, Pinkner JS, Chorell E, Dodson KW, Shaffer CL, Conover MS, Livny J, Hadjifrangiskou M, Almqvist F, Hultgren SJ - MBio (2014)

Global transcriptional and proteomic responses to ec240. (A) Genes whose expression is dysregulated by growth in ec240 rather than DMSO, as identified by RNA-Seq analysis. Genes were classified by using KEGG (http://www.genome.jp/kegg/) and EcoCyc (http://www.ecocyc.org). The genes in each functional category are shown as percentages of the total genes dysregulated. (B) MA plots of UTI89 gene abundance following treatment with ec240. Each dot represents an annotated UTI89 gene with the log2 of its relative abundance in ec240 versus that in DMSO (M) plotted against the average log2 of its abundance under both conditions (A). M and A values are based on data from three biological replicates under each growth condition. The dotted lines indicate 3-fold changes. (C) Fold changes in CUP gene transcripts altered in UTI89 grown with ec240 rather than the DMSO vehicle control, as determined by RNA-Seq. Genes are shown in their order within the operon. (D) Proteins dysregulated after growth in ec240 rather than DMSO, as determined by iTRAQ analysis. Proteins were classified by using KEGG (http://www.genome.jp/kegg/) and EcoCyc (http://www.ecocyc.org). The proteins in each functional category are shown as percentages of the total proteins dysregulated and include proteins with increased and decreased abundances. (E) Fold changes in CUP proteins altered in UTI89 grown with ec240 rather than the DMSO vehicle control, as determined by iTRAQ analysis. (F) Dysregulation by ec240 of siderophore-related and iron transport genes by both RNA-Seq and iTRAQ analysis. Shown are fold changes in transcript or protein levels in UTI89 grown in ec240 rather than DMSO. Genes are shown in their order within the operon.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4217179&req=5

fig2: Global transcriptional and proteomic responses to ec240. (A) Genes whose expression is dysregulated by growth in ec240 rather than DMSO, as identified by RNA-Seq analysis. Genes were classified by using KEGG (http://www.genome.jp/kegg/) and EcoCyc (http://www.ecocyc.org). The genes in each functional category are shown as percentages of the total genes dysregulated. (B) MA plots of UTI89 gene abundance following treatment with ec240. Each dot represents an annotated UTI89 gene with the log2 of its relative abundance in ec240 versus that in DMSO (M) plotted against the average log2 of its abundance under both conditions (A). M and A values are based on data from three biological replicates under each growth condition. The dotted lines indicate 3-fold changes. (C) Fold changes in CUP gene transcripts altered in UTI89 grown with ec240 rather than the DMSO vehicle control, as determined by RNA-Seq. Genes are shown in their order within the operon. (D) Proteins dysregulated after growth in ec240 rather than DMSO, as determined by iTRAQ analysis. Proteins were classified by using KEGG (http://www.genome.jp/kegg/) and EcoCyc (http://www.ecocyc.org). The proteins in each functional category are shown as percentages of the total proteins dysregulated and include proteins with increased and decreased abundances. (E) Fold changes in CUP proteins altered in UTI89 grown with ec240 rather than the DMSO vehicle control, as determined by iTRAQ analysis. (F) Dysregulation by ec240 of siderophore-related and iron transport genes by both RNA-Seq and iTRAQ analysis. Shown are fold changes in transcript or protein levels in UTI89 grown in ec240 rather than DMSO. Genes are shown in their order within the operon.
Mentions: We assessed the transcriptional profile of UTI89 grown with 250 µM ec240 or a dimethyl sulfoxide (DMSO) vehicle control to investigate the global effects of ec240 on UPEC biology. We utilized a relatively high concentration of ec240 in order to detail all possible “off-target” effects. RNA-Seq was conducted on triplicate biological samples grown under type 1 pilus-inducing conditions. We identified 52 gene transcripts that were altered ≥3-fold when UTI89 was grown in ec240 rather than the DMSO vehicle control. Of these 52 genes, 15 had decreased transcript levels and 37 had increased transcript levels. The transcript levels of CUP pilus genes were affected by the pilicide ec240, as were the transcript levels of genes with a variety of different functions, including motility, siderophore synthesis and transport, metabolism, translation, gene regulation, and stress response (Fig. 2A). There were also some hypothetical genes with dysregulated transcript levels (Fig. 2A). As shown in the MA plot (a plot of the log2 of the ratio of abundances of each transcript between the two conditions [M] plotted against the average log2 of abundance of that transcript in both conditions [A]), the pap and sfa genes encoding P and S pili had the most upregulated transcript levels, while the fim and flg genes encoding type 1 pili and flagella had the most downregulated transcript levels (Fig. 2B), with fim transcripts downregulated 8- to 17-fold, pap transcripts upregulated 4- to 49-fold, and sfa transcripts upregulated 4- to 25-fold (Fig. 2C). Furthermore, CUP operon genes cumulatively make up 46% of the genes dysregulated by >3-fold but 76% of the genes dysregulated by >10-fold after growth in ec240. This indicates that under type 1 pilus-inducing conditions, the major effect of ec240 is the dysregulation of CUP pili.

Bottom Line: Furthermore, the effect of ec240 on motility was abolished in the absence of the SfaB, PapB, SfaX, and PapX regulators.In contrast to the effects of ec240, deletion of the type 1 pilus operon led to increased S and P piliation and motility.Thus, ec240 dysregulated several uropathogenic Escherichia coli (UPEC) virulence factors through different mechanisms and independent of its effects on type 1 pilus biogenesis and may have potential as an antivirulence compound.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Microbiology and Microbial Pathogenesis and Center for Women's Infectious Disease Research, Washington University School of Medicine in St. Louis, St. Louis, Missouri, USA.

Show MeSH
Related in: MedlinePlus