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Identification Et Caractérisation D'arn Régulateurs Impliqués Dans La Identification et caractérisation d’ARN régulateurs impliqués dans la réponse au stress et la virulence chez Enterococcus faecalis Marine Salze To cite this version: Marine Salze. Identification et caractérisation d’ARN régulateurs impliqués dans la réponse au stress et la virulence chez Enterococcus faecalis. Santé. Normandie Université, 2019. Français. NNT : 2019NORMC403. tel-03221792 HAL Id: tel-03221792 https://tel.archives-ouvertes.fr/tel-03221792 Submitted on 10 May 2021 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THÈSE Pour obtenir le diplôme de doctorat Spécialité ASPECTS MOLECULAIRES ET CELLULAIRES DE LA BIOLOGIE Préparée au sein de l'Université de Caen Normandie Ιdentificatiοn et caractérisatiοn d'ΑRΝ régulateurs impliqués dans la répοnse au stress et la virulence chez Εnterοcοccus faecalis Présentée et soutenue par Marine SALZE Thèse soutenue publiquement le 09/05/2019 devant le jury composé de Mme HILDE DE REUSE Directeur de recherche, Institut Pasteur Rapporteur du jury Mme PASCALE ROMBY Directeur de recherche au CNRS, Université de Strasbourg Rapporteur du jury Mme NATHALIE CONNIL Maître de conférences HDR, Université Rouen Normandie Membre du jury Mme CECILE MULLER- PUJOL Maître de conférences, Université Caen Normandie Membre du jury M. HARALD PUTZER Directeur de recherche, Institut de biologie physico-chimique Président du jury M. ALAIN RINCE Professeur des universités, Université Caen Normandie Directeur de thèse Thèse dirigée par ALAIN RINCE, UR risques microbiens (Caen) Un scientifique dans son laboratoire est non seulement un technicien : il est aussi un enfant placé devant des phénomènes naturels qui l'impressionnent comme des contes de fées. - Marie Curie 1 Remerciements Je souhaite en premier lieu remercier le Docteur Hilde De Reuse et le Docteur Pascale Romby pour avoir accepté d’évaluer cette thèse et pour leur investissement en tant que rapporteurs. Je souhaiterais également remercier le Docteur Harald Putzer et le Docteur Nathalie Connil pour leur présence dans le jury et pour avoir accepté d’examiner mon travail de thèse. Je remercie également le Professeur Axel Hartke et le Professeur Alain Rincé qui m’ont accueillie au sein du laboratoire U2RM. Je tiens à remercier le Professeur Alain Rincé qui a dirigé ces travaux. Merci pour ta patience, pour le temps que tu as toujours réussi à me donner, et pour ta gentillesse. Et merci encore de m’avoir proposé ce sujet au moment où je m’étais résignée à ne pas faire de thèse. J’ai beaucoup appris à tes côtés, merci encore. Je souhaite adresser également tous mes remerciements au Docteur Cécile Muller pour avoir co-encadré cette thèse. Merci d’avoir guidé et soutenu pendant ces (plus de) trois ans la boule de stress que je suis, malgré les bévues et les chutes de confiance. Merci pour tout ce que tu m’as appris. Je te souhaite le meilleur pour la suite et bonne chance pour ton HDR. Je voudrais remercier toute l’équipe d’enseignants-chercheurs et de chercheurs du laboratoire, pour leur aide et leurs conseils en toute circonstance, mais aussi pour avoir donné le goût de la recherche à une petite étudiante de M1. Merci également à Benoit Bernay et Julien Pontin pour les manips de spectrométrie de masse. Merci également à Didier Goux pour les expériences de microscopie électronique. Merci aussi à Eliette Bisson, Nicolas Verneuil, Abdellah Benachour et Jean-Christophe Giard qui m’ont apporté leur aide précieuse pour les corrections de ce manuscrit. 2 Un merci tout particulièrement aux techniciennes, Isabelle et Anne-Cécile, que j’ai fait trimer à la fin de ma thèse dans la course aux derniers résultats. Merci pour votre aide indispensable. Je souhaite tous particulièrement remercier les thésards et ATER du laboratoire, Cindy, Loïc, Maxime (merci pour l’éthanol dans le tampon A4 !), Hakim, Delphine et Valentin. Merci pour les rires, le soutien, la Door of Shame Fame, les soirées, les jeux de mots nuls, les discussions éclairées et avisées du vendredi soir après 17h. Et surtout merci à tous pour cette atmosphère joyeuse qui a toujours régné autant que je me souvienne dans ce laboratoire, et qui a été un incroyable moteur pour la réussite de cette thèse. J’adresse une tendresse toute particulière à ma famille. Maman, Papa, merci pour le soutien, les sessions skype, les passages-éclairs. Tout cela n’avait pas de prix. Merci à mon frère Guillaume, soutien indéfectible malgré la distance, and thank you to my sister-in-law, Haruka, always with a nice word. Une tendresse pour mes magnifiques petites nièces, Lilwenn et Manon. Merci à Julien, Fiona, Magali, Di. Et surtout Camille. Ça fait 15 ans qu’on forme notre équipe de toquées, merci à toi d’être toujours là. Je vais terminer ces remerciements par Simon, qui a partagé ma vie et ma thèse malgré son éloignement, et continue encore. Tu es arrivé dans ma vie au milieu de mon doctorat, tu l’as chamboulée pour le mieux, merci pour tout. Merci à tous 3 Sommaire Liste des abréviations ........................................................................................................................... 5 Liste des figures .................................................................................................................................... 7 Liste des tableaux ................................................................................................................................. 9 Introduction ...................................................................................................................... 11 I. Cotexte de l’tude ........................................................................................................... 12 II. Le genre Enterococcus ...................................................................................................... 13 1. Historique ........................................................................................................... 13 2. Caractéristiques du genre Enterococcus ............................................................ 14 3. Enterococcus faecalis .......................................................................................... 15 a) Environnement et applications ............................................................................................. 15 b) Données génomiques ............................................................................................................ 16 c) Opportunisme et virulence ................................................................................................... 16 d) Métabolisme.......................................................................................................................... 18 III. La régulation transcriptionnelle et post-transcriptionnelle par les ARN régulateurs ............................................................................................... 21 1. Gralits sur la rgulatio de l’expressio des ges .................................... 21 2. Les petits ARN ..................................................................................................... 23 a) Les petits ARN codés en cis ................................................................................................... 23 b) Les petits ARN codés en trans ............................................................................................... 24 c) Cas particuliers ...................................................................................................................... 26 3. Les ARN liant les protéines ................................................................................. 26 4. Les CRISPR ........................................................................................................... 28 5. Les ’UTR UnTranslated Region) régulatrices ................................................... 28 a) Régulation par blocage ou déclenchement de la traduction ................................................ 29 b) Régulation par terminaison prématurée de la transcription ................................................ 30 6. Les ’UTR rgulatries......................................................................................... 31 IV. Les protéines impliquées dans le métabolisme des ARN .................... 32 1. Les protéines chaperonnes à ARN ...................................................................... 32 2. Le dégradosome ................................................................................................. 33 a) Les RNases ............................................................................................................................. 34 4 b) Les hélicases à motif DEAD .................................................................................................... 35 V. Les ARNs régulateurs chez E. faecalis ................................................. 37 VI. Contexte et objectifs .......................................................................... 41 Matériel et Méthodes ........................................................................................... 43 Résultats ...............................................................................................................................
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