Relationships between MDR proteins, bacteriocin production and proteolysis in Lactococcus lactis Gajic, Olivera

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1 University of Groningen Relationships between MDR proteins, bacteriocin production and proteolysis in Lactococcus lactis Gajic, Olivera IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 2003 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Gajic, O. (2003). Relationships between MDR proteins, bacteriocin production and proteolysis in Lactococcus lactis Groningen: s.n. Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date:

2 Relationships between MDR proteins, bacteriocin production and proteolysis in Lactococcus lactis Olivera Gaji

3 Cover : Fluorescence microscopic picture of Lactococcus lactis cells expressing LmrB fused with green fluorescence protein Printed by: Media Point, Leeuwarden This study was carried out at the Department of Genetics of the Groningen Biomolecular Sciences and and Biotechnology Institute (GBB) of the University of Groningen, the Netherlands. The research was financially supported by GBB.

4 RIJKSUNIVERSITEIT GRONINGEN Relationships between MDR proteins, bacteriocin production and proteolysis in Lactococcus lactis Proefschrift ter verkrijging van het doctoraat in de Wiskunde en Natuurwetenschappen aan de Rijksuniversiteit Groningen op gezag van de Rector Magnificus, dr. F. Zwarts, in het openbaar te verdedigen op vrijdag 24 januari 2003 om uur door Olivera Gaji geboren op 27 juni 1971 te Trstenik, Joegoslavië

5 Promotores: Co-promotor: Prof. dr. O.P. Kuipers Prof. dr. L. Topisirovi Dr. J. Kok Beoordelingscommissie: Prof. dr. I. Nes Prof. dr. G. Venema Prof. dr. W.N. Konings

6 Voorwoord Gotovo! The results of a few years of hard working summarised in a thesis. What was only meant to be a short visit in the summer of 98, but became a stay that lasted somewhat longer! The first acquaintance with the molecular genetics group was my contact with Gerard Venema. Although he was already retired when I started my work, he arranged a scholarship and some additional financial support afterwards. Gerard, hartelijk bedankt daarvoor. His successor, Oscar Kuipers, gave me the opportunity to work as an AIO in his group. Oscar, your enthusiasm for the multidrug resistance and bacteriocin work was motivating and constituted an important contribution to this thesis. Bedank voor al je vragen, suggesties en vertrouwen. I would also like to thank my second promotor, Ljubisa Topisirovi. Topi, naša saradnja je zapo ela još 95 godine kada sam po prvi put i osetila ari molekularne genetike i evo nastavila se, i rezultira ovom tezom. Hvala na razumevanju, savetima i dugim pismima diskutuju i o rezultatima. And Jan Kok, my co-promoter. Jan, thanks for the time and energy you spend on reading, discussing and polishing my manuscripts. Those sessions were highly teachable. I would also like to acknowledge the following persons that have contributed, in one way or another, to this thesis. Special thanks goes to Girbe for the inspiring supervision at the beginning of the project. Arie, Mozes and Peter, thanks for saving me a lot of time by washing glass, sterilising media and ordering all the (sometimes exotic) compounds. Emma, thanks for your help in filling out all the forms, which is rather difficult if you do not speak Dutch. And of course my colleagues that created a great atmosphere in the lab and at conferences. The Lactococci, my paranimf Anton (imam blot zato što imam seminar u ponedeljak, moja kultura je prljava and a lot more) Rober (Robertino, to my knowledge... ), Rasmus, Nathalie, Naomi, Wietske, Ana Rute, Chris, Richard, Harma, Sacha, Anne (H), Anne (de J), the ex-members student of mine Gertina (thanks for the help; chapter 2), Carmen, Patricia, Blandine, Christian, Mags, Michiel, Manolo, Mario, Giovanna, Tamara, Agnieszka, Kees and Jan Willem. Bacilli, thanks, especially the two French ladies Caroline and Elise. At the microbiology department I got a lot of support from my Spanish friend Abelardo, in introducing me to the techniques that are needed to study MDR proteins and his nice tortilla s, and Gerrit and Piotr, who I could always importune with my questions. I also thank Airi Palva and her group for collaboration. Although they did not contribute scientifically, family (in-laws) and friends played an important role behind the scenes. Mama, tata i Oliver, pregurasmo i ovo. Hvala za podršku, razumevanje i ljubav. Ni ne znate koliko mi to zna i. Oliver, ovo je jedinstvena prilika da te vidimo u fraku. Znam da eš dobro odigrati tu ulogu. And last but not least, Arjen and Dunja. Arjen, I do not know which language to use to thank you for everything you did. Ti i Dunja ste nešto najlepše što mi se desilo.

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8 Contents Chapter Introduction. Chapter Novel mechanism of bacteriocin secretion and immunity involving lactococcal MDR proteins. Chapter Antimicrobial peptides transported by bacterial MDR proteins and human P-glycoprotein. Chapter Dual regulation of lactococcin production by proteolytic action of the cell wall proteinase PrtP and by medium dependent transcriptional control. Chapter Proteinase and maturase gene expression in Lactococcus lactis is regulated by the transcriptional regulator CodY. Chapter Repetition of the cell wall spanning domain of lactococcal proteinase enhances cell sedimentation and interaction with eukaryotic cells. Chapter Summary and concluding remarks. Samenvatting en conclusies. Chapter Za neupu ene. Chapter Reference list.

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