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Dorado Morales, Pedro

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https://academica-e.unavarra.es/handle/2454/48915

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Dorado Morales

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Pedro

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Producción Agraria

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0000-0001-5760-1999

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811141

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2020 - 20213
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Subject: Staphylococcus aureus ×

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Now showing 1 - 3 of 3
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    PublicationOpen Access
    Systematic reconstruction of the complete two-component sensorial network in staphylococcus aureus
    (American Society for Microbiology, 2020) Rapún Araiz, Beatriz; Haag, Andreas F.; Gil Puig, Carmen; Dorado Morales, Pedro; Lasa Uzcudun, Íñigo; Ciencias de la Salud; Osasun Zientziak
    In bacteria, adaptation to changes in the environment is mainly controlled through two-component signal transduction systems (TCSs). Most bacteria contain dozens of TCSs, each of them responsible for sensing a different range of signals and controlling the expression of a repertoire of target genes (regulon). Over the years, identification of the regulon controlled by each individual TCS in different bacteria has been a recurrent question. However, limitations associated with the classical approaches used have left our knowledge far from complete. In this report, using a pioneering approach in which a strain devoid of the complete nonessential TCS network was systematically complemented with the constitutively active form of each response regulator, we have reconstituted the regulon of each TCS of S. aureus in the absence of interference between members of the family. Transcriptome sequencing (RNA-Seq) and proteomics allowed us to determine the size, complexity, and insulation of each regulon and to identify the genes regulated exclusively by one or many TCSs. This gain-of-function strategy provides the first description of the complete TCS regulon in a living cell, which we expect will be useful to understand the pathobiology of this important pathogen. IMPORTANCE Bacteria are able to sense environmental conditions and respond accordingly. Their sensorial system relies on pairs of sensory and regulatory proteins, known as two-component systems (TCSs). The majority of bacteria contain dozens of TCSs, each of them responsible for sensing and responding to a different range of signals. Traditionally, the function of each TCS has been determined by analyzing the changes in gene expression caused by the absence of individual TCSs. Here, we used a bacterial strain deprived of the complete TC sensorial system to introduce, one by one, the active form of every TCS. This gain-of-function strategy allowed us to identify the changes in gene expression conferred by each TCS without interference of other members of the family.
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    PublicationOpen 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 Zientziak
    Staphylococcus 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.
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    PublicationOpen Access
    Fitness cost evolution of natural plasmids of staphylococcus aureus
    (American Society for Microbiology, 2021) Dorado Morales, Pedro; Garcillán-Barcia, María Pilar; Lasa Uzcudun, Íñigo; Solano Goñi, Cristina; Ciencias de la Salud; Osasun Zientziak
    Plasmids have largely contributed to the spread of antimicrobial resistance genes among Staphylococcus strains. Knowledge about the fitness cost that plasmids confer on clinical staphylococcal isolates and the coevolutionary dynamics that drive plasmid maintenance is still scarce. In this study, we aimed to analyze the initial fitness cost of plasmids in the bacterial pathogen Staphylococcus aureus and the plasmid-host adaptations that occur over time. For that, we first designed a CRISPR (clustered regularly interspaced palindromic repeats)-based tool that enables the removal of native S. aureus plasmids and then transferred three different plasmids isolated from clinical S. aureus strains to the same-background clinical cured strain. One of the plasmids, pUR2940, obtained from a livestock-associated methicillin-resistant S. aureus (LA-MRSA) ST398 strain, imposed a significant fitness cost on both its native and the new host. Experimental evolution in a nonselective medium resulted in a high rate pUR2940 loss and selected for clones with an alleviated fitness cost in which compensatory adaptation occurred via deletion of a 12.8-kb plasmid fragment, contained between two ISSau10 insertion sequences and harboring several antimicrobial resistance genes. Overall, our results describe the relevance of plasmid-borne insertion sequences in plasmid rearrangement and maintenance and suggest the potential benefits of reducing the use of antibiotics both in animal and clinical settings for the loss of clinical multidrug resistance plasmids.