Evidences for the non-redundant function of A-type proteins ISCA1 and ISCA2 in iron-sulfur cluster biogenesis Lena Kristina Beilschmidt To cite this version: Lena Kristina Beilschmidt. Evidences for the non-redundant function of A-type proteins ISCA1 and ISCA2 in iron-sulfur cluster biogenesis. Organisation et fonctions cellulaires [q-bio.SC]. Université de Strasbourg, 2014. Français. NNT : 2014STRAJ031. tel-01297049 HAL Id: tel-01297049 https://tel.archives-ouvertes.fr/tel-01297049 Submitted on 2 Apr 2016 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. 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UNIVERSITÉ DE STRASBOURG Ecole doctorale des Sciences de la Vie et de la Santé Institut de Génetique et de Biologie Moléculaire et Cellulaire THÈSE présentée par : Lena Kristina BEILSCHMIDT soutenue le : 18 Novembre 2014 pour obtenir le grade de : Docteur de l’université de Strasbourg Discipline/ Spécialité : Aspects moléculaires et cellulaires de la biologie Evidences for the non-redundant function of A-type proteins ISCA1 and ISCA2 in iron-sulfur cluster biogenesis THÈSE dirigée par : Dr. PUCCIO Hélène (IGBMC, Illkirch) RAPPORTEURS : Dr. BALK Janneke (University of East Anglia, Norwich UK) Dr. BOUTON Cécile (Institut de Chimie des Substances Naturelles, Gif-sur-Yvette France) AUTRES MEMBRES DU JURY : Dr. OLLAGNIER-DE-CHOUDENS Sandrine (CEA, Grenoble France) Dr. CHARLET BERGUERAND Nicolas (IGBMC, Illkirch France) Acknowledgements To Lavrans, Miron and Arja Different people say different things at different times. What matters is what stays – what stays is what matters. 2 Acknowledgements I. Acknowledgements First of all, I want to thank the members of my PhD jury for reading and evaluating my thesis. Thank you for the time and the effort. I am indebted to Hélène Puccio for giving me the opportunity to do my PhD in her lab. Not only to be happy to “take me right away as a student” (as you said on the phone), but also for the stimulating surrounding for my research. Alain Martelli, I am indebted to you as well in many ways. I learned to decide and not always ask for your advice. Thanks for making me do my own mistakes and successes. Thank you for giving me the opportunity to teach and gaining the experience the CV is asking for. I am grateful for interacting with Marjorie Fournier for our collaboration concerning the MudPIT analysis, and thanks to Pascale Koebel who did all the AAV production. Further I want to thank all the common facilities that made scientific research possible. I want to thank all present and former lab members, too many to mention everyone in person. You contributed to the development of my project and the daily environment. I want to thank also many people from the department; especially Karim for being one of the few that showed scientific interest, gave support and honesty. A particular big thanks to the few but outstanding people that helped in solving each and every problem occurring in the interface between Lena – the German and the French administration; in particular the University of Strasbourg (inventing new troubles each year). Mainly these thanks go to Morgane, who made many phone calls for me, and I missed you for my last year, but also others (like Leila) helped. Anyone claiming that French speaking is not required for (scientific) life in the IGBMC should try it for one week. So, big thanks to each and every one that made the effort to make communication in english‐german‐french possible. I want to thank Ben and all present and former members of the SPB. The interactions, organizations and beers were lots of fun, success and an enriching experience. I want to thank Johann for all the interaction around teaching at the European school, and sharing beers over German football matches. There are always a few special ones to thank and I am afraid I will forget more than one person and aspect. So I want to apologize to the ones I missed. 3 Acknowledgements Leïla, you are one of the few that can discuss private life as a friend and still being critical and honest about science and a project over a coffee. I do hope for many future inter‐actions! We share ideas about fundamental science. Floriana, I want to thank you for being a true scientific friend ‐always critical but motivating to go on – in life and in science. Both of you enriched my life so much. Thanks to all friends that I found in the igbmc, many left the place before me. In particular Helena (the list, if it lay here, would be getting too long), Claire (always up for …), Nikolas (sarcastic from the beginning to the end), Serena (we will party together one day), Ana (thank you, seriously, thank you), Carlos (are you really from Sweden?), … and not to forget Mannu (the first coffee mate), Fred (sweetie always), Soumya (you know why) and Laurianne (Madeira, Paris, Berlin,…). In addition, I would like to mention Lana (Svetlana) Sirko that supported me in the moment of a very difficult decision and made sure that I can go my way. You not just always offered profound scientific discussion, but in this step your help was neither obvious nor self‐evident. Special thanks to ”mes coeurs” Anaïs and Mike. You made my life in Strasbourg a life ‐ a life outside of science, the institute and you provided family to me. Thanks for introducing me to your friends and your family, integrating the “silent‐german only speaking english” of the beginning and keeping the “funny‐sentence‐creator” from the end. You started the running and dinners, showing how to speak and to cook the French way. This also accounts for “mes cheris” Anaïs and Jean‐Christophe and Cyril – merci beaucoup. Jeanne, you became a special friend almost a granny. Thanks to all my other friends for being friends and all supportive actions during these last years, also here too many to mention them all now. Thank you! Danke! Merci! Gracias! Obrigada og Tusen Takk! Last but not least… will ich mich bei meiner Familie bedanken für all die Dinge, für die man eigentlich gar nicht danke sagen kann, weil sie wie selbstverständlich gegeben scheinen. Doch sie sind nicht selbstverständlich und trotzdem fällt es schwer, sie in Worte zu fassen. Danke für alles, jeden Moment, jede Hilfe und jeden Rat und jede Tat. Danke für die guten, aber auch die sprungfixen Gene, die ich mir wenigstens mit Arne teilen durfte. Danke Ihr drei! 4 Table of contents II. Table of contents Acknowledgements……………………………………………………………………………………………………………………… 3 Table of contents…………………………………………………………………………………………………………………………..5 List of Figures ........................................................................................................................................... 9 List of Manuscript Figures…….……..……………………………………………………………………………………………………10 List of Tables………………………….…………………………………………………………………………………….…………………..10 Preambule ………………………………………………………………………………………………………………………………..11 1. Introduction …………………………………………………………………………………………………….………………………12 1.1 Fe‐S cluster ................................................................................................................................ 12 1.2 Principle of Fe‐S biogenesis ....................................................................................................... 14 1.3 Essential nature of Fe‐S cluster and proteins ............................................................................ 16 1.3.1 Fe‐S cluster in DNA metabolism ........................................................................................ 17 1.3.2 Fe‐S cluster in the respiratory chain .................................................................................. 18 1.3.3 Fe‐S cluster in lipoic acid biosynthesis .............................................................................. 23 1.4 Fe‐ S biogenesis ......................................................................................................................... 25 1.4.1 Nascent cluster formation in mitochondria – the core machinery ................................... 26 1.4.2 Targeting mitochondrial Fe‐S proteins .............................................................................. 27 1.4.2.1 NFU1 and BOLA3 ‐ specific mitochondrial targeting factors ......................................... 27 1.4.2.2 IND1 protein required for in complex I formation ........................................................ 28 1.4.2.3 GLRX5 protein, a specific or general targeting factor? ................................................. 28 1.4.2.4 Conclusion on mitochondrial Fe‐S targeting ................................................................. 29 1.4.2.5 Cluster targeting from ISCU via HSCB to complex II: a new mechanism? ..................... 29 1.4.3 Extra‐mitochondrial Fe‐S clusters: Export and cytosolic Fe‐S assembly ........................... 30 1.5 The A‐type proteins and IBA57 ................................................................................................. 32 5 Table of contents 1.5.1 Bacterial A‐type proteins in Fe‐S biogenesis ..................................................................... 32 1.5.1.1 Fe‐S cluster binding versus Fe binding