Overexpressed proteins in hypervirulent clade 8 and clade 6 strains of Escherichia coli O157:H7 compared to E. coli O157:H7 EDL933 clade 3 strain
Escherichia coli O157:H7 is responsible for severe diarrhea and hemolytic uremic syndrome (HUS), and predominantly affects children under 5 years. The major virulence traits are Shiga toxins, necessary to develop HUS and the Type III Secretion System (T3SS) through which bacteria translocate effecto...
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| Format: | info:eu-repo/semantics/article |
| Language: | Inglés |
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2017
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| Online Access: | http://hdl.handle.net/20.500.12123/1115 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0166883 https://doi.org/10.1371/journal.pone.0166883 |
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| author | Amigo, Natalia Qi, Zhang Amadio, Ariel Qunjie, Zhang Marques da Silva, Wanderson Baiyuan, Cui Zhongjian, Chen Larzabal, Mariano Cataldi, Angel Adrian Jinlong, Bei |
| author_browse | Amadio, Ariel Amigo, Natalia Baiyuan, Cui Cataldi, Angel Adrian Jinlong, Bei Larzabal, Mariano Marques da Silva, Wanderson Qi, Zhang Qunjie, Zhang Zhongjian, Chen |
| author_facet | Amigo, Natalia Qi, Zhang Amadio, Ariel Qunjie, Zhang Marques da Silva, Wanderson Baiyuan, Cui Zhongjian, Chen Larzabal, Mariano Cataldi, Angel Adrian Jinlong, Bei |
| author_sort | Amigo, Natalia |
| collection | INTA Digital |
| description | Escherichia coli O157:H7 is responsible for severe diarrhea and hemolytic uremic syndrome (HUS), and predominantly affects children under 5 years. The major virulence traits are Shiga toxins, necessary to develop HUS and the Type III Secretion System (T3SS) through which bacteria translocate effector proteins directly into the host cell. By SNPs typing, E. coli O157:H7 was separated into nine different clades. Clade 8 and clade 6 strains were more frequently associated with severe disease and HUS. In this study, we aimed to identify differentially expressed proteins in two strains of E. coli O157:H7 (clade 8 and clade 6), obtained from cattle and compared them with the well characterized reference EDL933 strain (clade 3). Clade 8 and clade 6 strains show enhanced pathogenicity in a mouse model and virulence-related properties. Proteins were extracted and analyzed using the TMT-6plex labeling strategy associated with two dimensional liquid chromatography and mass spectrometry in tandem. We detected 2241 proteins in the cell extract and 1787 proteins in the culture supernatants. Attention was focused on the proteins related to virulence,
overexpressed in clade 6 and 8 strains compared to EDL933 strain. The proteins relevant overexpressed in clade 8 strain were the curli protein CsgC, a transcriptional activator (PchE), phage proteins, Stx2, FlgM and FlgD, a dienelactone hydrolase, CheW and CheY, and the SPATE protease EspP. For clade 6 strain, a high overexpression of phage proteins was detected, mostly from Stx2 encoding phage, including Stx2, flagellin and the protease TagA, EDL933_p0016, dienelactone hydrolase, and Haemolysin A, amongst others with
unknown function. Some of these proteins were analyzed by RT-qPCR to corroborate the proteomic data. Clade 6 and clade 8 strains showed enhanced transcription of 10 out of 12 genes compared to EDL933. These results may provide new insights in E. coli O157:H7 mechanisms of pathogenesis. |
| format | info:eu-repo/semantics/article |
| id | INTA1115 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | INTA11152017-12-01T14:14:17Z Overexpressed proteins in hypervirulent clade 8 and clade 6 strains of Escherichia coli O157:H7 compared to E. coli O157:H7 EDL933 clade 3 strain Amigo, Natalia Qi, Zhang Amadio, Ariel Qunjie, Zhang Marques da Silva, Wanderson Baiyuan, Cui Zhongjian, Chen Larzabal, Mariano Cataldi, Angel Adrian Jinlong, Bei Genética Enfermedades de los Animales Escherichia Coli Genetics Animal Diseases Escherichia coli O157:H7 is responsible for severe diarrhea and hemolytic uremic syndrome (HUS), and predominantly affects children under 5 years. The major virulence traits are Shiga toxins, necessary to develop HUS and the Type III Secretion System (T3SS) through which bacteria translocate effector proteins directly into the host cell. By SNPs typing, E. coli O157:H7 was separated into nine different clades. Clade 8 and clade 6 strains were more frequently associated with severe disease and HUS. In this study, we aimed to identify differentially expressed proteins in two strains of E. coli O157:H7 (clade 8 and clade 6), obtained from cattle and compared them with the well characterized reference EDL933 strain (clade 3). Clade 8 and clade 6 strains show enhanced pathogenicity in a mouse model and virulence-related properties. Proteins were extracted and analyzed using the TMT-6plex labeling strategy associated with two dimensional liquid chromatography and mass spectrometry in tandem. We detected 2241 proteins in the cell extract and 1787 proteins in the culture supernatants. Attention was focused on the proteins related to virulence, overexpressed in clade 6 and 8 strains compared to EDL933 strain. The proteins relevant overexpressed in clade 8 strain were the curli protein CsgC, a transcriptional activator (PchE), phage proteins, Stx2, FlgM and FlgD, a dienelactone hydrolase, CheW and CheY, and the SPATE protease EspP. For clade 6 strain, a high overexpression of phage proteins was detected, mostly from Stx2 encoding phage, including Stx2, flagellin and the protease TagA, EDL933_p0016, dienelactone hydrolase, and Haemolysin A, amongst others with unknown function. Some of these proteins were analyzed by RT-qPCR to corroborate the proteomic data. Clade 6 and clade 8 strains showed enhanced transcription of 10 out of 12 genes compared to EDL933. These results may provide new insights in E. coli O157:H7 mechanisms of pathogenesis. Inst. de Biotecnología Fil: Amigo, Natalia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Qi, Zhang. Guangdong ‘Academy of Agricultural Sciences. AGRO-Biological Gene Research Center; China Fil: Amadio, Ariel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina Fil: Qunjie, Zhang. Guangdong ‘Academy of Agricultural Sciences. AGRO-Biological Gene Research Center; China Fil: Marques da Silva, Wanderson. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Zhongjian, Chen. Guangdong ‘Academy of Agricultural Sciences. AGRO-Biological Gene Research Center; China Fil: Larzabal, Mariano. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Jinlong, Bei. Guangdong ‘Academy of Agricultural Sciences. AGRO-Biological Gene Research Center; China Fil: Cataldi, Angel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina 2017-09-04T13:41:40Z 2017-09-04T13:41:40Z 2016 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/1115 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0166883 Amigo N, Zhang Q, Amadio A, Zhang Q, Silva WM, Cui B, et al. (2016) Overexpressed Proteins in Hypervirulent Clade 8 and Clade 6 Strains of Escherichia coli O157:H7 Compared to E. coli O157:H7 EDL933 Clade 3 Strain. PLoS ONE 11(11): e0166883 1932-6203 (Online) https://doi.org/10.1371/journal.pone.0166883 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf PLoS ONE 11 (11) : e016688 (2016) |
| spellingShingle | Genética Enfermedades de los Animales Escherichia Coli Genetics Animal Diseases Amigo, Natalia Qi, Zhang Amadio, Ariel Qunjie, Zhang Marques da Silva, Wanderson Baiyuan, Cui Zhongjian, Chen Larzabal, Mariano Cataldi, Angel Adrian Jinlong, Bei Overexpressed proteins in hypervirulent clade 8 and clade 6 strains of Escherichia coli O157:H7 compared to E. coli O157:H7 EDL933 clade 3 strain |
| title | Overexpressed proteins in hypervirulent clade 8 and clade 6 strains of Escherichia coli O157:H7 compared to E. coli O157:H7 EDL933 clade 3 strain |
| title_full | Overexpressed proteins in hypervirulent clade 8 and clade 6 strains of Escherichia coli O157:H7 compared to E. coli O157:H7 EDL933 clade 3 strain |
| title_fullStr | Overexpressed proteins in hypervirulent clade 8 and clade 6 strains of Escherichia coli O157:H7 compared to E. coli O157:H7 EDL933 clade 3 strain |
| title_full_unstemmed | Overexpressed proteins in hypervirulent clade 8 and clade 6 strains of Escherichia coli O157:H7 compared to E. coli O157:H7 EDL933 clade 3 strain |
| title_short | Overexpressed proteins in hypervirulent clade 8 and clade 6 strains of Escherichia coli O157:H7 compared to E. coli O157:H7 EDL933 clade 3 strain |
| title_sort | overexpressed proteins in hypervirulent clade 8 and clade 6 strains of escherichia coli o157 h7 compared to e coli o157 h7 edl933 clade 3 strain |
| topic | Genética Enfermedades de los Animales Escherichia Coli Genetics Animal Diseases |
| url | http://hdl.handle.net/20.500.12123/1115 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0166883 https://doi.org/10.1371/journal.pone.0166883 |
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