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Identifier 000441540
Title Investigation for the detection of putative intracellular targets of Shigella
Alternative Title Μελέτη για την ανίχνευση υποκυτταρικών στόχων βακτηρίων του γένους Shigella
Author Αραπίτσας, Νικόλαος Π.
Thesis advisor Σαρρής, Παναγιώτης
Reviewer Παυλίδης, Ιωάννης
Σκούλικα, Ευθυμία
Abstract Shigella is a dangerous primate-restricted pathogen strongly linked with worldwide diarrhea-associated mortality. Its growing development of antibiotic resistance and the lack of an effective vaccine constitutes a health issue, especially for developing countries. Shigella infects its hosts by injecting a suite of effector proteins into the human colonic epithelial cells, causing the shigellosis disease. A better understanding of the Shigella pathogenicity will aid the fight against shigellosis. Despite Shigella being a human pathogen, it recently became evident that it can infect and proliferate in Arabidopsis thaliana, employing effectors used for pathogenesis in humans. That indicates that Shigella interacts with both a human and a plant host. In this thesis, we sought to elucidate and highlight the presence of common strategies and principles governing the host-pathogen interactions between plants and animals. In a cross-kingdom approach, we attempted to check whether the integrated decoys of the cytoplasmic nucleotide-binding leucine-rich repeat (NLR) receptors of the plant innate immune system can recognize and interact with the effector proteins of the highly specialized human pathogen Shigella. We employed a yeast two-hybrid system in which twelve Shigella effector proteins were individually coexpressed with each of twenty-three plant NLR integrated decoys (IDs) in yeast cells (Saccharomyces cerevisiae). With the yeast two-hybrid screenings, we tried to detect interactions between these effector proteins and IDs. The transformation of yeast cells with the Shigella effector proteins showed that OspF, VirA, IpgD, IpaJ, and OspC1 are toxic upon expression in yeast leading to growth inhibition. A mild autoactivation ability was observed during the yeast two-hybrid screenings for the EXO70 ID and a stronger one for the WRKY_Barley of the Hordeum vulgare. Interestingly, a putative interaction was identified between the effector protein IpaJ and the WRKY_Barley ID. The study of the interplay of plant proteins with the proteins from animal pathogens could improve the understanding of the virulence strategies that animal pathogens employ. Furthermore, this framework can highlight novel targets for the prevention and treatment of the diseases that human pathogens cause. In future experiments, we will further study the IpaJ – WRKY_Barley pair to validate the presence of such an interaction and analyze the putative underlying molecular mechanism. Furthermore, we will focus on Shigella effector proteins that are homologous to effector proteins from plant pathogens, which have known plant protein targets. Finally, we aim to investigate whether such plant proteins can interact with the Shigella effectors.
Language English
Subject Host-pathogen interaction
Innate immunity
Yeast-two-hybrid screening
Αλληλεπίδραση παθογόνου ξενιστή
Δοκιμασία δύο υβριδίων
Issue date 2021-07-30
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
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