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; Zhāng, Qí; Amadío, Ariel Fernando; Zhang, Qun‐Jie; Silva, Wanderson Marques da; Cui, Baiyuan; Chen, Zhongjian; Larzábal, Mariano; Bei, Jinlong; Cataldi, Angel

doi:10.1371/journal.pone.0166883

Cited by 14 publications

(16 citation statements)

References 73 publications

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“…A study in 2008 identified a subtype of E. coli O157:H7, termed clade 8, associated with increased risk of HUS [10]. Although numerous studies have investigated virulence factor expression that may be responsible for this association [11–14], studies confirming the association have been limited, suggesting an effect of varying strength and specificity [15–17]. …”

Section: Introductionmentioning

confidence: 99%

Importance of case age in the purported association between phylogenetics and hemolytic uremic syndrome inEscherichia coliO157:H7 infections

Tarr¹,

Shringi

Oltean

et al. 2018

Epidemiol. Infect.

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Escherichia coli O157:H7 is the largest cause of hemolytic uremic syndrome (HUS). Previous studies proposed that HUS risk varies across the E. coli O157:H7 phylogenetic tree (hypervirulent clade 8), but the role of age in the association is unknown. We determined phylogenetic lineage of E. coli O157:H7 isolates from 1160 culture-confirmed E. coli O157:H7 cases reported in Washington State, 2004-2015. Using generalised estimating equations, we tested the association between phylogenetic lineage and HUS. Age was evaluated as an effect modifier. Among 1082 E. coli O157:H7 cases with both phylogenetic lineage and HUS status (HUS n = 76), stratified analysis suggested effect modification by age. Lineages IIa and IIb, relative to Ib, did not appear associated with HUS in children 0-9-years-old. For cases 10-59-years-old, lineages IIa and IIb appeared to confer increased risk of HUS, relative to lineage Ib. The association reversed in ⩾60-year-olds. Results were similar for clade 8. Phylogenetic lineage appears to be associated with HUS risk only among those ⩾10-years-old. Among children <10, the age group most frequently affected, lineage does not explain progression to HUS. However, lineage frequency varied across age groups, suggesting differences in exposure and/or early disease manifestation.

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Section: Introductionmentioning

confidence: 99%

Importance of case age in the purported association between phylogenetics and hemolytic uremic syndrome inEscherichia coliO157:H7 infections

Tarr¹,

Shringi

Oltean

et al. 2018

Epidemiol. Infect.

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“…The EHEC O157:H7 strains Rafaela II (clade 8) and 7.1 Anguil (clade 6) isolated from cattle in Argentina and EDL933 (clade 3) strain recovered from a patient in USA were routinely maintained in Luria-Bertani broth (LB, Difco Laboratories, USA) or in LB 1.5% bacteriological agar plates, at 37°C. For the proteomic studies, bacterial strains were cultured as previously described by Amigo et al (7). Overnight cultures of the different EHEC O157:H7 strains growth in LB were inoculated (1:50) in Dulbecco’s modified Eagle’s medium (DMEM)-F12 nutrient until reach the mid-exponential growth phase (OD 600nm =0.6) under a 5% CO 2 atmosphere at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…After bacterial growth, protein extractions were performed according to Amigo et al (7). Three biological replicates of each culture were centrifuged at 5000 × g for 20 min at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…In a previous study, we reported the use of isobaric tags for comparative quantitation (TMT) method to identify the differentially expressed proteins among three EHEC O157:H7 isolates: Rafaela II (Clade 8), Anguil 7.1 (Clade 6) and EDL933 (Clade 3) (7). The proteome differences observed among these strains are related mainly to proteins involved in both virulence and cellular metabolism; which might reflect the virulence potential of each strain (7). The aim of the present study was to promote a more comprehensive insight into the molecular basis of EHEC O157:H7 physiology and pathogenesis.…”

Section: Introductionmentioning

confidence: 99%

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Quantification of EnterohemorrhagicEscherichia coliO157:H7 proteome using TMT-Based Analysis

Bei

Amigo

et al. 2018

Preprint

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25Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a human pathogen responsible for 26 diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS). EHEC infection is 27 distributed worldwide and numerous outbreaks of diseases caused by enterohemorrhagic 28 have been reported. To promote a comprehensive insight into the molecular basis of EHEC 29 O157:H7 physiology and pathogenesis, the combined proteome of EHEC O157:H7 strains, 30 Clade 8 and Clade 6 isolated from cattle in Argentina, and the standard EDL933 (clade 3) 31 strain has been analyzed. TMT (Tandem Mass Tags)-based quantitative proteomic and 32 emPAI analyses were performed to estimate the protein abundance in EHEC proteome. 33 2,234 non-redundant proteins of EHEC O157:H7 were identified. A comparison of this 34 result with in silico data of EHEC O157:H7 genome showed that approximately 40% of the 35 predicted proteome of this pathogen were covered. According to the emPAI analysis, 85 36 proteins were among the most abundant (e.g. GAPDH, FliC H-antigen, Enolase, and 37 GroEL). Tellurite resistance proteins were also highly abundant. COG analysis showed that 38 although most of the identified proteins are related to cellular metabolism, the majority of 39 the most abundant proteins are associated with translation processes. A KEGG enrichment 40 analysis revealed that Glycolysis / Gluconeogenesis was the most significant pathway. On 41 the other hand, the less abundant detected proteins are those related to DNA processes, cell 42 respiration and prophage. Among the proteins that composed the Type III Secretion 43 System, the most abundant protein was EspA. Altogether, the results show a subset of 44 important proteins that contribute to physiology and pathogenicity of EHEC O157:H7. 45 IMPORTANCE 46 3 The study of the abundance of proteins present within a complex mixture of proteins in a 47 cell, under different conditions, can provide important information about the activities of 48 individual protein components and protein networks that are cornerstones for the 49 comprehension of physiological adaptations in response to biological demands promoted 50 by environmental changes. We generated a comprehensive and accurate quantitative list of 51 EHEC O157:H7 proteome, which provides a description of the most abundant proteins 52 produced by this pathogen that were related to physiology and pathogenesis of EHEC. This 53 study provides information and extends the understanding on functional genomics and the 54 biology of this pathogen.55 Keywords: EHEC, Tandem mass tags (TMT), protein quantitation, bacterial proteomic 56 57 58 Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a zoonotic pathogen belonging to 59 Shiga toxin-producing E. coli (STEC) and responsible for different diseases as diarrhea, 60 hemorrhagic colitis and hemolytic uremic syndrome (HUS). HUS is distributed worldwide 61 and considered to be a public health problem in several countries (1,2). Unfortunately, 62 Argentina is the country with the highest incidence of HUS i...

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“…Reversed-Phase Fractionation. The mixed TMT labelled peptides were desalted with Sep-Pak SPE cartridges (Waters) for subsequent hpRP separation in a Dionex Ulti-Mate 3000 model as described previously [10]. Finally, 48 collected fractions were merged into 12 mixtures (Supplementary figure 1).…”

Section: High Phmentioning

confidence: 99%

A Phenotypic Characterization of Two Isolates of a Multidrug-Resistant Outbreak Strain of Mycobacterium tuberculosis with Opposite Epidemiological Fitness

Bei

Bigi

Lima

et al. 2020

BioMed Research International

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Tuberculosis (TB) is an infectious disease, caused by Mycobacterium tuberculosis, primarily affecting the lungs. The M. tuberculosis strain of the Haarlem family named M was responsible for a large multidrug-resistant TB (MDR-TB) outbreak in Buenos Aires. This outbreak started in the early 1990s and in the mid 2000s still accounted for 29% of all MDR-TB cases in Argentina. By contrast, a clonal variant of strain M, named 410, has caused a single tuberculosis case since the onset of the outbreak. The molecular bases of the high epidemiological fitness of the M strain remain unclear. To assess its unique molecular properties, herein, we performed a comparative protein and lipid analysis of a representative clone of the M strain (Mp) and the nonprosperous M variant 410. We also evaluated their growth in low pH. The variant 410 had higher levels of latency proteins under standard conditions and delayed growth at low pH, suggesting that it is more sensitive to stress stimuli than Mp. Moreover, Mp showed higher levels of mycolic acids covalently attached to the cell wall and lower accumulation of free mycolic acids in the outer layer than the 410 strain. The low expression of latency proteins together with the reduced content of surface mycolic acids may facilitate Mp to evade the host immune responses.

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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

Cited by 14 publications

References 73 publications

Importance of case age in the purported association between phylogenetics and hemolytic uremic syndrome inEscherichia coliO157:H7 infections

Importance of case age in the purported association between phylogenetics and hemolytic uremic syndrome inEscherichia coliO157:H7 infections

Quantification of EnterohemorrhagicEscherichia coliO157:H7 proteome using TMT-Based Analysis

A Phenotypic Characterization of Two Isolates of a Multidrug-Resistant Outbreak Strain of Mycobacterium tuberculosis with Opposite Epidemiological Fitness

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