1. Deep impact of the inactivation of the SecA2-only protein export pathway on the proteosurfaceome of Listeria monocytogenes
- Author
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Pilar Teixeira, Magdalena Popowska, Christophe Chambon, Mickaël Desvaux, Michel Hébraud, Morgan Guilbaud, Rafał Ostrowski, Jean-Marie Herry, Ingrid Chafsey, Nelly Caccia, Marie-Noëlle Bellon-Fontaine, Joana Azeredo, Microbiologie Environnement Digestif Santé (MEDIS), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Clermont Auvergne (UCA), University of Warsaw (UW), Paris-Saclay Food and Bioproduct Engineering (SayFood), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University of Minho [Braga], Plateforme Exploration du Métabolisme-Composante Protéomique (PFEM-CP), Qualité des Produits Animaux (QuaPA), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Universidade do Minho
- Subjects
Proteomics ,Cell surface properties ,Cellular differentiation ,[SDV]Life Sciences [q-bio] ,Biophysics ,Virulence ,Protein Export Pathway ,medicine.disease_cause ,Secretomic analysis ,Biochemistry ,Bacterial secretion systems ,03 medical and health sciences ,Listeria monocytogenes ,Bacterial Proteins ,Biotecnologia Médica [Ciências Médicas] ,medicine ,SecA2-export pathway ,Humans ,030304 developmental biology ,Adenosine Triphosphatases ,0303 health sciences ,Science & Technology ,030306 microbiology ,Chemistry ,Biofilm ,Membrane Transport Proteins ,Cell biology ,Secretory protein ,Cell surface proteome ,Ciências Médicas::Biotecnologia Médica ,Cell envelope ,Biogenesis - Abstract
Listeria monocytogenes presents a dimorphism associated to the SecA2 activity with cells having a normal rod shape or a dysmorphic elongated filamentous form. Besides variation of the cell and colony morphotype, this cell differentiation has profound ecophysiological and physiopathological implications with collateral effects on virulence and pathogenicity, biotope colonisation, bacterial adhesion and biofilm formation. This suggests the SecA2-only protein export could influence the listerial cell surface, which was investigated first by characterising its properties in L. monocytogenes wt and secA2. The degree of hydrophilicity and Lewis acid-base properties appeared significantly affected upon SecA2 inactivation. As modification of electrostatic properties would owe to modification in the composition of cell-surface proteins, the proteosurfaceome was further investigated by shotgun label-free proteomic analysis with a comparative relative quantitative approach. Following secretomic analysis, the protein secretion routes of the identified proteins were mapped considering the cognate transport and post-translocational maturation systems, as well as protein categories and subcellular localisation. Differential protein abundance profiles coupled to network analysis revealed the SecA2 dependence of 48 proteins, including some related to cell envelope biogenesis, translation and protein export, which could account for modifications of adhesion and cell properties of L. monocytogenes upon SecA2 inactivation. This investigation unravelled the profound influence of SecA2 activity on the cell surface properties and proteosurfaceome of L. monocytogenes, which provides advanced insights about its ecophysiopathology. Significance L. monocytogenes is a foodborne zoonotic pathogen and etiological agent of human listeriosis. This species presents a cellular dimorphism associated to the SecA2 activity that has profound physiopathological and ecophysiological implications with collateral effects on bacterial virulence and colonisation. To explore the influence of the SecA2-only protein export on the listerial cell, the surface properties of L. monocytogenes expressing or depleted of SecA2 was characterised by microelectrophoresis, microbial affinity to solvents and contact angles analyses. As modifications of hydrophilicity and Lewis acid-base electrostatic properties would owe to modification in the composition of cell-surface proteins, the proteinaceous subset of the surfaceome, i.e. the proteosurfaceome, was investigated further by shotgun label-free proteomic analysis. This subproteome appeared quite impacted upon SecA2 inactivation with the identification of proteins accounting for modifications in the cell surface properties. The profound influence of SecA2 activity on the cell surface of L. monocytogenes was unravelled, which provides advanced insights about its ecophysiopathology., INRAE (“Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement”, previously called INRA, “Institut National de la Recherche Agronomique”), NCN (“Narodowe Centrum Nauki”) National Science Centre Poland (n°2013/09/B/NZ6/00710), ANR (“Agence National de la Recherche”) PathoFood project (n°ANR-17-CE21-0002), COST (European Cooperation in Science and Technology) Action FA1202 BacFoodNet, RMT (“Réseau Mixte Technologique”) CHLEAN (“Conception Hygiénique des Lignes et Equipements et Amélioration de la Nettoyabilité pour une alimentation saine et sure”) and France-Poland CampusFrance EGIDE PHC (“Programme Hubert Curien”) POLONIUM 2013 (n°28298ZE), info:eu-repo/semantics/publishedVersion
- Published
- 2022