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The Venom of Nasonia Vitripennis: Involvement in the Host-Parasitoid Interaction and Putative Biomedical Application

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The Venom of Nasonia Vitripennis: Involvement in the Host-Parasitoid Interaction and Putative Biomedical Application The venom of Nasonia vitripennis: involvement in the host-parasitoid interaction and putative biomedical application Ellen Danneels Promotor: Prof. Dr. Dirk de Graaf Co-promotors: Prof. Dr. Kathleen Van Craenenbroeck Em. Prof. Dr. Frans Jacobs Academic year 2014-2015 Thesis submitted in fulfilment of the requirements for the degree of Doctor in Sciences: Biochemistry and Biotechnology Department of Biochemistry and Microbiology The research described in this thesis was performed at the Laboratory of Molecular Entomology and Bee Pathology, Ghent University. This work was partly funded by the Research Foundation-Flanders (FWO) (G041708N). Copyright © Ellen Danneels. All rights reserved. No parts of this work may be reproduced; any quotations must acknowledge this source. Supervisors Promotor: Prof. Dr. Dirk de Graaf Laboratory of Molecular Entomology and Bee Pathology (L-MEB) Department of Biochemistry and Microbiology Faculty of Science, Ghent University, Belgium Co-promotor: Prof. Dr. Kathleen Van Craenenbroeck Laboratory of Eukaryotic Gene Expression and Signal Transduction (LEGEST) Department of Physiology Faculty of Science, Ghent University, Belgium Co-promotor: Em. Prof. Dr. Frans Jacobs Honeybee Valley Faculty of Sciences, Ghent University, Belgium Examination committee Prof. Dr. Savvas Savvides (Ghent University, Belgium, chairman) Prof. Dr. Dirk de Graaf (Ghent University, Belgium, secretary) Prof. Dr. Kathleen Van Craenenbroeck (Ghent University, Belgium) Em. Prof. Dr. Frans Jacobs (Ghent University, Belgium) Prof. Dr. Karolien De Bosscher (VIB, Ghent University, Belgium)* Prof. Dr. Claude Libert (VIB, Ghent University, Belgium)* Dr. Eveline Verhulst (Wageningen University, The Netherlands)* Prof. Dr. Roger Huybrechts (KU Leuven, Belgium)* * Members of the reading committee Dean: Prof. Dr. Herwig Dejonghe Rector: Prof. Dr. Anne De Paepe Please refer to this work as: Danneels, E. L. (2014) The venom of Nasonia vitripennis: involvement in the parasitoid-host interaction and putative biomedical application. PhD thesis, Ghent University. Author’s email address: [email protected] DANKWOORD Tijdens mijn studies heb ik altijd beweerd dat doctoreren niet voor mij was weggelegd… Ik was zogezegd genetisch voorbestemd om in het onderwijs te gaan en ging prompt na mijn studies in Leuven die richting uit. Maar toen ik tijdens een interim in een middelbare school in Brugge de vacature voorgeschoteld kreeg voor een assistentschap, en dan nog wel in Gent, dichter bij huis dus, lonkte een ambitieuzere academische start waar ik met veel leergierigheid ben op ingestapt. De combinatie onderzoek en onderwijs bleek voor mij de perfecte combo en al snel raakte ik gebeten (of beter gestoken) door de bijen. Twee jaar lang kreeg ik de kans om mee te draaien in lopende bijenonderzoekingen en betrokken te worden in een samenwerking met goede mede-ex-studenten uit de KUL. Maar stilaan groeide mijn fascinatie voor een kleiner insect die recent zijn intrede had gemaakt in het labo: Nasonia vitripennis. Ik kreeg de keuze om een eventuele switch te maken in doctoraatsonderwerp en heb resoluut gekozen om voor de sprong in het diepe te gaan: het gif van Nasonia vitripennis. Het opzetten van de kweek en naderhand ook van zijn gastheer, de welriekende vleesvliegen, verliepen niet altijd zonder slag of stoot. Maar ondertussen draaien beide kweken routineus, al moet ik zeggen dat de geur van rottende lever nooit zal wennen. Vijf jaar lang heb ik me vastgebeten in de boeiende interactie tussen beide insecten en ik moet zeggen, het heeft me geen seconde verveeld. Met heel veel weemoed kijk ik terug op 7 jaar werken als assistent aan de UGent, een job die me enorm veel voldoening en plezier heeft bezorgd. De belangrijkste persoon in de mede-realisatie van dit werk is zonder twijfel mijn promotor Prof. Dirk de Graaf. Je enthousiasme in het onderzoek en excellente begeleiding, maken van jou de beste promotor die je je als startend doctoraatstudent kunt wensen. Door je niet- aflatende vertrouwen en steun in zowel mijn onderzoeksexperimenten als mijn onderwijstaken, ben ik als doctoraatstudente steeds kunnen blijven groeien en verbeteren. Naast de dagelijkse werkomgeving hebben we ook veel lief en leed gedeeld. In die 7 jaar hebben we van te veel mensen definitief afscheid moeten nemen en ook daarbij had ik het gevoel dat ik steeds bij je terecht kon. Maar laat ons vooral ook de plezante momenten niet vergeten, zoals de pizza-for-papers, de barbecues en de vele gezellige koffiepauzes. Enorm bedankt voor alles! Maar oorspronkelijk ben ik begonnen met mijn doctoraat bij DE bijenman van België, Prof. Frans Jacobs. Op de momenten dat het op het werk wat minder ging, wist je me er altijd aan te herinneren dat het belangrijkste uiteindelijk familie en gezin zijn, en ik geef je volledig gelijk! Bedankt voor het vertrouwen en de kansen die je me hebt gegeven. Twee is altijd beter dan één. De betrekking van een tweede labo met een enorme expertise bij de onderzoekingen in het tweede luik van dit proefschrift, is daarom een gigantische meerwaarde geweest. Bedankt Prof. Kathleen Van Craenenbroeck voor het gastvrij openstellen van je labo en de aangename samenwerking. Daarnaast zou ik ook alle overige leden van de examencommissie en de begeleidingscommissie willen bedanken voor hun terechte opmerkingen en hun inspanning om bij dit proefschrift de puntjes op de i te zetten. Ik heb het geluk gehad dat ik voor het aanleren van de RNAi techniek 4 weken heb kunnen vertoeven in het Centre for Ecological and Evolutionary Studies aan de Rijksuniversiteit Groningen. De warme gastvrije ontvangst en perfecte ondersteuning stonden haaks tegenover de ijzige kille -7°C die er heerste toen ik er verbleef. Bedankt Prof. Leo Beukeboom, Prof. Louis van de Zande, Prof. Bregje Wertheim, Eveline Verhulst, Anna Rensink, Elzemiek Geuverink, Sylvia Gerritsma en Rogier Houwerzijl voor de fijne periode! Maar het grootste deel van mijn doctoraat werd ik perfect omringd door een grote groep zeer behulpzame collega’s waar ik met onderzoeksproblemen, maar vooral ook met mijn eigen verhaal steeds bij terecht kon. Een speciale merci aan Marleen en Lina voor jullie raad en daad en onze gezellige babbels over ons kiendjes! Maar ook aan Matthias, Ellen, Jorgen, Tine, Sara, Karel, Karen, Bea, Linde, Prof. Sarah Gerlo, Jolien, An, Kamila en Nadia bedankt voor de hulp bij de kweken (zowel van insecten als van cellen), het luisterend oor als mijn experimenten maar weer eens mislukt waren, maar ook de ontspannende zwempartijtjes tussendoor en de vele gezellige koffiepauzes! Een collega die veel voor me heeft betekend, maar die jammergenoeg dit dankwoord niet meer kan nalezen, is Dieter. Ik ben enorm dankbaar dat ik je heb mogen leren kennen en had dit moment ook graag met jou gedeeld. Ik hoop dat je rust mag vinden en we komen elkaar nog wel tegen. Clairette, heel erg bedankt voor onze vele babbels waardoor we Dieter toch nog een beetje onder ons houden! Jammergenoeg wat veraf, maar toch ook steeds betrokken en begaan, zijn mijn goeie West- Vlaamse vrienden, nu weliswaar over heel België verspreid, maar toch houden we nog geregeld contact. We zaten allemaal in hetzelfde schuitje en fier mag ik nu ook als laatste in de rij jullie uitnodigen op de doctoraatsverdediging. Jaja, KULAK still rules! Uiteraard waren ook mijn ouders, ‘kleine’ zus, schoonbroer Stijn en schoonfamilie van groot belang bij het welslagen van dit doctoraat. De keuze om biologie te gaan studeren kwam misschien niet zo onverwacht, maar de beslissing om uiteindelijk te gaan doctoreren was dat wellicht iets meer. De meerdere pogingen om toch nog maar eens te proberen begrijpen wat het nu eigenlijk is wat ik hele dagen zit te doen in Gent en de niet-aflatende steun en fierheid voor ‘de (schoon)dochter/(schoon)zus aan de universiteit’, heeft me altijd ontroerd. Heel erg merci! Helemaal op het einde komen natuurlijk mijn twee allergrootste schatten. Zonder David was ik waarschijnlijk zelfs nooit begonnen met doctoreren. Jouw oneindige geduld en onvoorwaardelijke steun hebben me altijd het volle vertrouwen gegeven om de dingen met twee handen vast te grijpen. Je hebt me nooit iets in de weg gestaan, in tegendeel, je hebt me altijd gepusht om het beste in mezelf naar boven te halen. En daar ben ik je oneindig dankbaar voor! Samen hebben we al heel wat opgebouwd en bereikt. Maar het allermooiste is toch wel ons kleine poppemie, Emmelien. Hoewel de vele slapeloze nachten me vaak met kleine oogjes naar het werk hebben doen vertrekken, als ze je ’s avonds terug in de armen komt aangelopen, vergeet je weer volledig alle zorgen. Mijn klein bolleke, je beseft het misschien nog niet, maar je overvloed aan energie maakt telkens opnieuw mijn dag weer goed. Dantjewel keppemie! TABLE OF CONTENTS List of abbreviations......………………………………………………………………………………………………... 1 Summary............................................................................................................................. 3 Samenvatting...................................................................................................................... 5 PART I. INTRODUCTION Chapter 1. General introduction......................................................................................... 9 1. Parasitoids: the ‘Trojan insects’...................................................................................... 10 2. Tiny but mighty: Nasonia vitripennis.............................................................................. 12 2.1. Life cycle..........................................................................................................
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