Evaluation of CpxRA as a therapeutic target for uropathogenic Escherichia coli infections

Lana Dbeibo, Julia J. van Rensburg, Sara N. Smith, Kate R. Fortney, Dharanesh Gangaiah, Hongyu Gao, Juan Marzoa, Yunlong Liu, Harry L.T. Mobley, Stanley Spinola

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

CpxRA is an envelope stress response system found in all members of the family Enterobacteriaceae; CpxA has kinase activity for CpxR and phosphatase activity for phospho-CpxR, a transcription factor. CpxR also accepts phosphate groups from acetyl phosphate, a glucose metabolite. Activation of CpxR increases the transcription of genes encoding membrane repair and downregulates virulence determinants. We hypothesized that activation of CpxR could serve as an antimicrobial/antivirulence strategy and discovered compounds that activate CpxR in Escherichia coli by inhibiting CpxA phosphatase activity. As a prelude to testing such compounds in vivo, here we constructed cpxA (in the presence of glucose, CpxR is activated because of a lack of CpxA phosphatase) and cpxR (system absent) deletion mutants of uropathogenic E. coli (UPEC) CFT073. By RNA sequencing, few transcriptional differences were noted between the cpxR mutant and its parent, but in the cpxA mutant, several UPEC virulence determinants were downregulated, including the fim and pap operons, and it exhibited reduced mannose-sensitive hemagglutination of guinea pig red blood cells in vitro. In competition experiments with mice, both mutants were less fit than the parent in the urine, bladder, and kidney; these fitness defects were complemented in trans. Unexpectedly, in single-strain challenges, only the cpxA mutant was attenuated for virulence in the kidney but not in the bladder or urine. For the cpxA mutant, this may be due to the preferential use of amino acids over glucose as a carbon source in the bladder and urine by UPEC. These studies suggest that CpxA phosphatase inhibitors may have some utility for treating complex urinary tract infections.

Original languageEnglish (US)
Article numbere00798-17
JournalInfection and Immunity
Volume86
Issue number3
DOIs
StatePublished - Mar 1 2018

Fingerprint

Uropathogenic Escherichia coli
Escherichia coli Infections
Phosphoric Monoester Hydrolases
Virulence
Urinary Bladder
Urine
Glucose
Down-Regulation
RNA Sequence Analysis
Kidney
Hemagglutination
Enterobacteriaceae
Therapeutics
Operon
Mannose
Urinary Tract Infections
Guinea Pigs
Transcription Factors
Phosphotransferases
Carbon

Keywords

  • Antivirulence
  • CpxA
  • CpxR
  • CpxRA
  • Escherichia coli
  • UPEC
  • Uropathogenic
  • Virulence determinants

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

Cite this

Dbeibo, L., van Rensburg, J. J., Smith, S. N., Fortney, K. R., Gangaiah, D., Gao, H., ... Spinola, S. (2018). Evaluation of CpxRA as a therapeutic target for uropathogenic Escherichia coli infections. Infection and Immunity, 86(3), [e00798-17]. https://doi.org/10.1128/IAI.00798-17

Evaluation of CpxRA as a therapeutic target for uropathogenic Escherichia coli infections. / Dbeibo, Lana; van Rensburg, Julia J.; Smith, Sara N.; Fortney, Kate R.; Gangaiah, Dharanesh; Gao, Hongyu; Marzoa, Juan; Liu, Yunlong; Mobley, Harry L.T.; Spinola, Stanley.

In: Infection and Immunity, Vol. 86, No. 3, e00798-17, 01.03.2018.

Research output: Contribution to journalArticle

Dbeibo, L, van Rensburg, JJ, Smith, SN, Fortney, KR, Gangaiah, D, Gao, H, Marzoa, J, Liu, Y, Mobley, HLT & Spinola, S 2018, 'Evaluation of CpxRA as a therapeutic target for uropathogenic Escherichia coli infections', Infection and Immunity, vol. 86, no. 3, e00798-17. https://doi.org/10.1128/IAI.00798-17
Dbeibo L, van Rensburg JJ, Smith SN, Fortney KR, Gangaiah D, Gao H et al. Evaluation of CpxRA as a therapeutic target for uropathogenic Escherichia coli infections. Infection and Immunity. 2018 Mar 1;86(3). e00798-17. https://doi.org/10.1128/IAI.00798-17
Dbeibo, Lana ; van Rensburg, Julia J. ; Smith, Sara N. ; Fortney, Kate R. ; Gangaiah, Dharanesh ; Gao, Hongyu ; Marzoa, Juan ; Liu, Yunlong ; Mobley, Harry L.T. ; Spinola, Stanley. / Evaluation of CpxRA as a therapeutic target for uropathogenic Escherichia coli infections. In: Infection and Immunity. 2018 ; Vol. 86, No. 3.
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abstract = "CpxRA is an envelope stress response system found in all members of the family Enterobacteriaceae; CpxA has kinase activity for CpxR and phosphatase activity for phospho-CpxR, a transcription factor. CpxR also accepts phosphate groups from acetyl phosphate, a glucose metabolite. Activation of CpxR increases the transcription of genes encoding membrane repair and downregulates virulence determinants. We hypothesized that activation of CpxR could serve as an antimicrobial/antivirulence strategy and discovered compounds that activate CpxR in Escherichia coli by inhibiting CpxA phosphatase activity. As a prelude to testing such compounds in vivo, here we constructed cpxA (in the presence of glucose, CpxR is activated because of a lack of CpxA phosphatase) and cpxR (system absent) deletion mutants of uropathogenic E. coli (UPEC) CFT073. By RNA sequencing, few transcriptional differences were noted between the cpxR mutant and its parent, but in the cpxA mutant, several UPEC virulence determinants were downregulated, including the fim and pap operons, and it exhibited reduced mannose-sensitive hemagglutination of guinea pig red blood cells in vitro. In competition experiments with mice, both mutants were less fit than the parent in the urine, bladder, and kidney; these fitness defects were complemented in trans. Unexpectedly, in single-strain challenges, only the cpxA mutant was attenuated for virulence in the kidney but not in the bladder or urine. For the cpxA mutant, this may be due to the preferential use of amino acids over glucose as a carbon source in the bladder and urine by UPEC. These studies suggest that CpxA phosphatase inhibitors may have some utility for treating complex urinary tract infections.",
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