cAMP Receptor Protein Positively Regulates the Expression of Genes Involved in the Biosynthesis of Klebsiella oxytoca Tilivalline Cytotoxin
Klebsiella oxytoca is a resident of the human gut. However, certain K. oxytoca toxigenic strains exist that secrete the nonribosomal peptide tilivalline (TV) cytotoxin. TV is a pyrrolobenzodiazepine that causes antibiotic-associated hemorrhagic colitis (AAHC). The biosynthesis of TV is driven by enz...
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| Format: | Article |
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Frontiers Media S.A.
2021-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.743594/full |
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| author | Diana Rodríguez-Valverde Diana Rodríguez-Valverde Nancy León-Montes Nancy León-Montes Jorge Soria-Bustos Jessica Martínez-Cruz Jessica Martínez-Cruz Ricardo González-Ugalde Ricardo González-Ugalde Sandra Rivera-Gutiérrez Jorge A. González-y-Merchand Roberto Rosales-Reyes Lázaro García-Morales Hidetada Hirakawa James G. Fox Jorge A. Girón Miguel A. De la Cruz Miguel A. Ares Miguel A. Ares |
| author_facet | Diana Rodríguez-Valverde Diana Rodríguez-Valverde Nancy León-Montes Nancy León-Montes Jorge Soria-Bustos Jessica Martínez-Cruz Jessica Martínez-Cruz Ricardo González-Ugalde Ricardo González-Ugalde Sandra Rivera-Gutiérrez Jorge A. González-y-Merchand Roberto Rosales-Reyes Lázaro García-Morales Hidetada Hirakawa James G. Fox Jorge A. Girón Miguel A. De la Cruz Miguel A. Ares Miguel A. Ares |
| author_sort | Diana Rodríguez-Valverde |
| collection | DOAJ |
| description | Klebsiella oxytoca is a resident of the human gut. However, certain K. oxytoca toxigenic strains exist that secrete the nonribosomal peptide tilivalline (TV) cytotoxin. TV is a pyrrolobenzodiazepine that causes antibiotic-associated hemorrhagic colitis (AAHC). The biosynthesis of TV is driven by enzymes encoded by the aroX and NRPS operons. In this study, we determined the effect of environmental signals such as carbon sources, osmolarity, and divalent cations on the transcription of both TV biosynthetic operons. Gene expression was enhanced when bacteria were cultivated in tryptone lactose broth. Glucose, high osmolarity, and depletion of calcium and magnesium diminished gene expression, whereas glycerol increased transcription of both TV biosynthetic operons. The cAMP receptor protein (CRP) is a major transcriptional regulator in bacteria that plays a key role in metabolic regulation. To investigate the role of CRP on the cytotoxicity of K. oxytoca, we compared levels of expression of TV biosynthetic operons and synthesis of TV in wild-type strain MIT 09-7231 and a Δcrp isogenic mutant. In summary, we found that CRP directly activates the transcription of the aroX and NRPS operons and that the absence of CRP reduced cytotoxicity of K. oxytoca on HeLa cells, due to a significant reduction in TV production. This study highlights the importance of the CRP protein in the regulation of virulence genes in enteric bacteria and broadens our knowledge on the regulatory mechanisms of the TV cytotoxin. |
| first_indexed | 2024-12-19T16:29:29Z |
| format | Article |
| id | doaj.art-d12904ea35924fa5beaec01db707beb0 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-19T16:29:29Z |
| publishDate | 2021-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-d12904ea35924fa5beaec01db707beb02022-12-21T20:14:15ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-09-011210.3389/fmicb.2021.743594743594cAMP Receptor Protein Positively Regulates the Expression of Genes Involved in the Biosynthesis of Klebsiella oxytoca Tilivalline CytotoxinDiana Rodríguez-Valverde0Diana Rodríguez-Valverde1Nancy León-Montes2Nancy León-Montes3Jorge Soria-Bustos4Jessica Martínez-Cruz5Jessica Martínez-Cruz6Ricardo González-Ugalde7Ricardo González-Ugalde8Sandra Rivera-Gutiérrez9Jorge A. González-y-Merchand10Roberto Rosales-Reyes11Lázaro García-Morales12Hidetada Hirakawa13James G. Fox14Jorge A. Girón15Miguel A. De la Cruz16Miguel A. Ares17Miguel A. Ares18Unidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Mexico City, MexicoDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, MexicoUnidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Mexico City, MexicoDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, MexicoUnidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Mexico City, MexicoUnidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Mexico City, MexicoDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, MexicoUnidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Mexico City, MexicoDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, MexicoDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, MexicoDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, MexicoUnidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City, MexicoDepartamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Mexico City, MexicoDepartment of Bacteriology, Gunma University Graduate School of Medicine, Maebashi, JapanDivision of Comparative Medicine, Massachusetts Institute of Technology, Cambridge, MA, United StatesCentro de Detección Biomolecular, Benemérita Universidad Autónoma de Puebla, Puebla, MexicoUnidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Mexico City, MexicoUnidad de Investigación Médica en Enfermedades Infecciosas y Parasitarias, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Mexico City, MexicoDepartamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, MexicoKlebsiella oxytoca is a resident of the human gut. However, certain K. oxytoca toxigenic strains exist that secrete the nonribosomal peptide tilivalline (TV) cytotoxin. TV is a pyrrolobenzodiazepine that causes antibiotic-associated hemorrhagic colitis (AAHC). The biosynthesis of TV is driven by enzymes encoded by the aroX and NRPS operons. In this study, we determined the effect of environmental signals such as carbon sources, osmolarity, and divalent cations on the transcription of both TV biosynthetic operons. Gene expression was enhanced when bacteria were cultivated in tryptone lactose broth. Glucose, high osmolarity, and depletion of calcium and magnesium diminished gene expression, whereas glycerol increased transcription of both TV biosynthetic operons. The cAMP receptor protein (CRP) is a major transcriptional regulator in bacteria that plays a key role in metabolic regulation. To investigate the role of CRP on the cytotoxicity of K. oxytoca, we compared levels of expression of TV biosynthetic operons and synthesis of TV in wild-type strain MIT 09-7231 and a Δcrp isogenic mutant. In summary, we found that CRP directly activates the transcription of the aroX and NRPS operons and that the absence of CRP reduced cytotoxicity of K. oxytoca on HeLa cells, due to a significant reduction in TV production. This study highlights the importance of the CRP protein in the regulation of virulence genes in enteric bacteria and broadens our knowledge on the regulatory mechanisms of the TV cytotoxin.https://www.frontiersin.org/articles/10.3389/fmicb.2021.743594/fullCRPtilivalline cytotoxinaroXnpsAKlebsiella oxytoca |
| spellingShingle | Diana Rodríguez-Valverde Diana Rodríguez-Valverde Nancy León-Montes Nancy León-Montes Jorge Soria-Bustos Jessica Martínez-Cruz Jessica Martínez-Cruz Ricardo González-Ugalde Ricardo González-Ugalde Sandra Rivera-Gutiérrez Jorge A. González-y-Merchand Roberto Rosales-Reyes Lázaro García-Morales Hidetada Hirakawa James G. Fox Jorge A. Girón Miguel A. De la Cruz Miguel A. Ares Miguel A. Ares cAMP Receptor Protein Positively Regulates the Expression of Genes Involved in the Biosynthesis of Klebsiella oxytoca Tilivalline Cytotoxin Frontiers in Microbiology CRP tilivalline cytotoxin aroX npsA Klebsiella oxytoca |
| title | cAMP Receptor Protein Positively Regulates the Expression of Genes Involved in the Biosynthesis of Klebsiella oxytoca Tilivalline Cytotoxin |
| title_full | cAMP Receptor Protein Positively Regulates the Expression of Genes Involved in the Biosynthesis of Klebsiella oxytoca Tilivalline Cytotoxin |
| title_fullStr | cAMP Receptor Protein Positively Regulates the Expression of Genes Involved in the Biosynthesis of Klebsiella oxytoca Tilivalline Cytotoxin |
| title_full_unstemmed | cAMP Receptor Protein Positively Regulates the Expression of Genes Involved in the Biosynthesis of Klebsiella oxytoca Tilivalline Cytotoxin |
| title_short | cAMP Receptor Protein Positively Regulates the Expression of Genes Involved in the Biosynthesis of Klebsiella oxytoca Tilivalline Cytotoxin |
| title_sort | camp receptor protein positively regulates the expression of genes involved in the biosynthesis of klebsiella oxytoca tilivalline cytotoxin |
| topic | CRP tilivalline cytotoxin aroX npsA Klebsiella oxytoca |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.743594/full |
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