Biology subjects

Larzabal, M.

Publications and source records attributed to Larzabal, M..

3 recordsLinked to original sources

bioRxiv2025

Global expression profile of Enterohemorrhagic Escherichia coli O157:H7 in phagosome of murine macrophages

EHEC O157:H7 is responsible for Hemolytic Uremic Syndrome (HUS) outbreaks in humans and its virulence is associated with Shiga toxin (Stx). During the pathogenesis of HUS, EHEC O157:H7 is phagocytized by the intestinal host macrophage into the phagosome. According to our study, a low percentage of phagocytized bacteria survived in the murine macrophage phagosome. To understand the initials mechanisms involved in bacterial persistence, we performed a global profile of bacterial RNAseq analysis in the early phagosome of murine macrophage and in vitro assays with more extreme stress conditions. The defense strategy of an early murine phagosome consisted on producing DNA damage, membrane damage, acid pH and nitric oxide (NO) agents. These hostile conditions triggering the bacterial SOS response, lipid biosynthesis and NO detoxification enzymes in response to DNA damage, membrane damage and nitric oxide agents, respectively. In turn, to survive, the bacterium conserves energy by downregulating flagellar biosynthesis, T3SS and T6SS virulence mechanisms. In contrast, stx2a expression is upregulated. At the same time, it increases the ribosomal levels and amino acid synthesis to react more effectively against adverse conditions. Under more extreme stress conditions, EHEC O157:H7 expressed genes related to an acidic environment (pH 4.5) upregulating the acid stress response pathways AR2-5, whereas, upon high concentrations of hydrogen peroxide, it transcribed higher levels of genes of the OxyR pathway and subsequently hydrogen peroxide oxidative stress-related genes.

microbiology↗

bioRxiv2024

Antibacterial T6SS1 cluster identification and bioinformatics characterization of their putative effectors in Shiga toxin-producing Escherichia coli

Shiga toxing-producing Escherichia coli (STEC) O22:H8 strain is a serotype occasionally isolated in Argentinian cattle. Preliminary works showed that the cattle carrying STEC O22:H8 strains could not be experimentally colonized by EHEC O157:H7 strain. The type 6 secretion system (T6SS) is one of the most versatile virulence mechanisms involved in delivering effectors, particularly the T6SS1 translocate antibacterial toxins effectors during bacterial competition for niche-space. In this work, we could evidence the molecular bases of the success of STEC O22:H8 (154) strain during bacterial competition against EHEC O157:H7 strains. The genome sequence of STEC O22:H8 (154) allowed us to identify a complete T6SS1 cluster. In addition, we identify and characterized several putative T6SS1-antibacterial effectors encoded inside the T6SS1 clusters and in genomic pathogenic islands. Competition assays against EHEC O157:H7 strain confirmed the antibacterial activity of STEC O22:H8 (154) strain in vitro. Considering the absent of T6SS1 in STEC strains, we proposed the recent horizontal transfer acquisition and the most probably donor belong to the same Escherichia coli species. A safe STEC O22:H8 (154){Delta} stx would be used as a new strategy to fight STECs in bovine intestinal colonization, leading in a reduction in beef contamination and consequently HUS cases in humans.

microbiology↗

bioRxiv2018

Quantification of Enterohemorrhagic Escherichia coli O157:H7 proteome using TMT-Based Analysis

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a human pathogen responsible for diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS). EHEC infection is distributed worldwide and numerous outbreaks of diseases caused by enterohemorrhagic have been reported. To promote a comprehensive insight into the molecular basis of EHEC O157:H7 physiology and pathogenesis, the combined proteome of EHEC O157:H7 strains, Clade 8 and Clade 6 isolated from cattle in Argentina, and the standard EDL933 (clade 3) strain has been analyzed. TMT (Tandem Mass Tags)-based quantitative proteomic and emPAI analyses were performed to estimate the protein abundance in EHEC proteome. 2,234 non-redundant proteins of EHEC O157:H7 were identified. A comparison of this result with in silico data of EHEC O157:H7 genome showed that approximately 40% of the predicted proteome of this pathogen were covered. According to the emPAI analysis, 85 proteins were among the most abundant (e.g. GAPDH, FliC H-antigen, Enolase, and GroEL). Tellurite resistance proteins were also highly abundant. COG analysis showed that although most of the identified proteins are related to cellular metabolism, the majority of the most abundant proteins are associated with translation processes. A KEGG enrichment analysis revealed that Glycolysis / Gluconeogenesis was the most significant pathway. On the other hand, the less abundant detected proteins are those related to DNA processes, cell respiration and prophage. Among the proteins that composed the Type III Secretion System, the most abundant protein was EspA. Altogether, the results show a subset of important proteins that contribute to physiology and pathogenicity of EHEC O157:H7.\n\nIMPORTANCEThe study of the abundance of proteins present within a complex mixture of proteins in a cell, under different conditions, can provide important information about the activities of individual protein components and protein networks that are cornerstones for the comprehension of physiological adaptations in response to biological demands promoted by environmental changes. We generated a comprehensive and accurate quantitative list of EHEC O157:H7 proteome, which provides a description of the most abundant proteins produced by this pathogen that were related to physiology and pathogenesis of EHEC. This study provides information and extends the understanding on functional genomics and the biology of this pathogen.

microbiology↗