Publication
Quantitative proteome analysis of an Escherichia coli exposed to tetracycline reveals multiple affected cytoplasmic metabolic processes
| dc.contributor.author | Jones-Dias, Daniela | |
| dc.contributor.author | Carvalho, Ana Sofia | |
| dc.contributor.author | Moura, Inês Barata | |
| dc.contributor.author | Dill, Brian | |
| dc.contributor.author | Molina, Henrik | |
| dc.contributor.author | Manageiro, Vera | |
| dc.contributor.author | Caniça, Manuela | |
| dc.contributor.author | Matthiesen, Rune | |
| dc.date.accessioned | 2017-02-16T16:28:49Z | |
| dc.date.available | 2017-02-16T16:28:49Z | |
| dc.date.issued | 2016-04 | |
| dc.description.abstract | Background: Many studies have already focused on the evaluation of variations in protein expression caused by antibiotic exposure in tetracycline resistant microorganisms. However, little is known about the metabolic response of genetically resistant bacterial populations to antibiotic exposure. In this study we used a liquid chromatography-mass spectrometry-based (LC-MS/MS) proteomics to evaluate the global cytoplasmic metabolic changes of a resistant Escherichia coli isolate, when challenged with tetracycline. Material/methods: IsolateEcAamb278 was recovered from a soil sample, used for the intensive farming of tomato plants and showed nonsusceptibility to all categories of β-lactams except carbapanems and cefoxitin, and tetracycline. Previous genomic characterization revealed the presence of TEM-1, CTX-M-1, Sul2,TetA and TetB resistance determinants. Single colonies of E. coliEcAamb278 were used to extract crude cytoplasmic protein fraction using non detergent cell lysis buffer. The proteomic profile of E. coli EcAmb278 was determined by label-free LC-MS/MS upon tetracycline stress in a comparative study, using as referencethe proteome of the same strain nonexposed to antibiotics. The samples were analyzed by nano LC-MS/MS, using a Q-Exactive mass spectrometer. All data was searched with VEMS. Proteins were quantified by spectral counting and mziXIC, followed by iBAQ estimation. Results: The complete proteome yielded about 1484 proteins, using a 1% FDR as cut off. The comparison of the proteome profiles of the two E. coli EcAamb278 samples pointed to several proteins with altered expression under tetracycline stress conditions. The twelve most significant (FDRcorrected p<0.05) proteins differentially regulated by more than two-fold were involved in DNA metabolism, transcription, virulence, intracellular trafficking and secretion, biosynthesis of vitamins, other processes, and unknown functions. For instance, (1) acyl-CoA thioester hydrolase, involved in biosynthesis of coenzyme A; (2) bacterial NAD+-dependent DNA ligase, plays a critical role in DNA replication, recombination and repair in all living organisms; and (3) adhesion protein FimH, which confirmed the pathogenicity of this environmental-borne E. coli. Overall, several other detected proteins were associated with the process of antibiotic resistance, virulence and acquisition and transfer of foreign DNA (AcrA, TolC, MdtE, Omps, TnsE, Colicin peptides, among others). Conclusions: These results indicate that E. coli responses to tetracycline are related to protein translation as well as metabolic regulation. We observed differentially regulation of several metabolic proteins though not belonging to the canonical antibiotic resistance pathway, suggesting an integrated metabolic bacteria response to antibiotic exposure, which can potentially be explored for drug development. | pt_PT |
| dc.description.version | info:eu-repo/semantics/publishedVersion | pt_PT |
| dc.identifier.citation | Jones-Dias, D., Carvalho, A. S., Moura, I. B., Dill, B. D., Molina, H., Manageiro, V., Caniça, M., Matthiesen, R. Quantitative proteome analysis of an Escherichia coli exposed to tetracycline reveals multiple affected cytoplasmic metabolic processes. 2016. 26th European Congress of Clinical Microbiology and Infectious Diseases, Amsterdam, Netherlands. P0770. | pt_PT |
| dc.identifier.uri | http://hdl.handle.net/10400.18/4218 | |
| dc.language.iso | eng | pt_PT |
| dc.peerreviewed | yes | pt_PT |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc/4.0/ | pt_PT |
| dc.subject | Antibacterial Resistance Mechanisms | pt_PT |
| dc.subject | Proteomics | pt_PT |
| dc.subject | Escherichia coli | pt_PT |
| dc.subject | Tetracycline Stress | pt_PT |
| dc.subject | Resistência aos Antimicrobianos | pt_PT |
| dc.title | Quantitative proteome analysis of an Escherichia coli exposed to tetracycline reveals multiple affected cytoplasmic metabolic processes | pt_PT |
| dc.type | conference object | |
| dspace.entity.type | Publication | |
| oaire.citation.conferencePlace | Amsterdam, Netherlands | pt_PT |
| oaire.citation.title | 26th European Congress of Clinical Microbiology and Infectious Diseases (ECCMID), 9-12 April 2016 | pt_PT |
| rcaap.rights | openAccess | pt_PT |
| rcaap.type | conferenceObject | pt_PT |