Dorado Morales, Pedro
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Dorado Morales
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Pedro
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Producción Agraria
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Publication Open Access The biofilm-associated surface protein Esp of Enterococcus faecalis forms amyloid-like fibers(Nature Research, 2020) Taglialegna, Agustina; Matilla Cuenca, Leticia; Dorado Morales, Pedro; Navarro, Susanna; Ventura, Salvador; Garnett, James A.; Lasa Uzcudun, Íñigo; Valle Turrillas, Jaione; Ciencias de la Salud; Osasun ZientziakFunctional amyloids are considered as common building block structures of the biofilm matrix in different bacteria. In previous work, we have shown that the staphylococcal surface protein Bap, a member of the Biofilm-Associated Proteins (BAP) family, is processed and the fragments containing the N-terminal region become aggregation-prone and self-assemble into amyloid-like structures. Here, we report that Esp, a Bap-orthologous protein produced by Enterococcus faecalis, displays a similar amyloidogenic behavior. We demonstrate that at acidic pH the N-terminal region of Esp forms aggregates with an amyloid-like conformation, as evidenced by biophysical analysis and the binding of protein aggregates to amyloid-indicative dyes. Expression of a chimeric protein, with its Esp N-terminal domain anchored to the cell wall through the R domain of clumping factor A, showed that the Esp N-terminal region is sufficient to confer multicellular behavior through the formation of an extracellular amyloid-like material. These results suggest that the mechanism of amyloid-like aggregation to build the biofilm matrix might be widespread among BAP-like proteins. This amyloid-based mechanism may not only have strong relevance for bacteria lifestyle but could also contribute to the amyloid burden to which the human physiology is potentially exposed.Publication Open Access Genetic tools derived from Staphylococcus aureus for biotechnological applications in Gram-positive bacteria(2021) Dorado Morales, Pedro; Lasa Uzcudun, Íñigo; Solano Goñi, Cristina; Ciencias de la Salud; Osasun ZientziakStaphylococcus aureus is a versatile human pathogen that has emerged as one of the most successful infectious agents of recent times, able to cause a range of diseases including skin and soft tissue infections, endocarditis, sepsis, pneumonia, osteomyelitis, bacteremia, and abscesses in organ tissues. Besides its clinical relevance, S. aureus has served as a model to study fundamental cellular processes, such as biofilm formation, the regulatory functions of small RNAs or growth and division of spherical cocci. Based on the accumulated knowledge of S. aureus biology, the availability of database resources and the advances in high-throughput genome sequencing, in this work we have aimed at developing new genetic tools derived from S. aureus for biotechnological applications in Gram-positive bacteria.