Structure and mechanism of the RNA polymerase II CTD phosphatase Scp1 and large-scale preparation of the RNA polymerase II-TFIIF complex [Elektronische Ressource] / Tomislav Kamenski

Tomislav Kamenski

Sujets

• Biologie

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2006
• Nombre de lectures: 30
• Langue: English
• Poids de l'ouvrage: 5 Mo

Extrait

Dissertation zur Erlangung des Doktorgrades der Fakultät für Chemie und
Pharmazie der Ludwig-Maximilians-Universität München








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Tomislav Kamenski
aus Zagreb, Kroatien
2006 Erklärung
Diese Dissertation wurde im Sinne von §13 Abs. 3 bzw. 4 der Promotionsordnung
vom 29. Januar 1998 von Herrn Prof. Dr. Patrick Cramer betreut.

Ehrenwörtliche Versicherung
Diese Dissertation wurde selbständig, ohne unerlaubte Hilfe erarbeitet.

München, den 2. Mai 2006







Dissertation eingereicht am 4. Mai 2006
1. Gutachter: Prof. Dr. Patrick Cramer
2. Gutachter: Prof. Dr. Ralf-Peter Jansen
Mündliche Prüfung am 29. Mai 2006 Acknowledgements
First of all I owe my gratitude to my PhD supervisor Prof. Patrick Cramer for providing
me excellent conditions for independent research work and for constant support.
I am most grateful to Claudia, for being a great companion on the long and difficult
way of TFIIF project and for loud laughs at the time when the project did not seem
promising. Toni Meinhart, thank you for being patient with me, while learning basics
of crystallography. I would also like to thank Stefan Benkert for great effort in
fermenting yeast, Marian Kalocay, for providing me with useful vectors and tips.
Karim, thank you for accompanying me to fast food restaurants (willingly) after long
days in the lab and Michela for good laughs and late evening jogging rounds. To
Hubert, Florian, Laurent and other lab members, I am grateful for being great
colleagues, always willing to help, and for nice atmosphere in the laboratory. I would
like to thank especially Kathrin Gmelch for being an excellent diploma student, as
well as Susanna Heilmeier for introducing me to phosphatase project. I am grateful to
Anass for great collaboration, exchange of scientific views and I wish him all the luck
in the continuation of the project.
I would like to thank Katja Sträßer and Heidi Feldmann for providing great advice and
being ready to teach me about yeast.
I would also like to thank members of my PhD thesis committee, Alexander Brehm
and Ralf-Peter Jansen for a lot of advice.
I wish to thank Ita Grui, for her expertise on RNA biochemistry, as well as Prof.
Ivana Weygand-uraševi for support even while being abroad.
I thank Boehringer Ingelheim Fonds for generous financial support and possibility to
meet other young motivated scientists with similar interests.
Finally, I am deeply indebted to my family for all their help and support during my
education.



Summary
1 Introduction
1.1 Eukaryotic RNA polymerases ...................................................................... 1
1.2 Structure of Pol II ......................................................................................... 2
1.3 Eukaryotic transcription cycle....................................................................... 3
1.4 TFIIF is a unique eukaryal general transcription factor ................................ 9
1.4.1 Roles of TFIIF during PIC formation and promoter escape .................... 13
1.4.2 Roles of TFIIF in elongation ................................................................... 14
1.4.3 TFIIF interactions with Pol II ................................................................... 14
1.5 Carboxy-terminal domain of Pol II (CTD) ................................................... 16
1.6 CTD phosphorylation and transcriptional regulation .................................. 17
1.7 CTD kinases .............................................................................................. 18
1.8 CTD phosphatase Ssu72........................................................................... 19
1.9 CTD phosphatases of the Fcp1 family ....................................................... 21
1.10 Aims of this work........................................................................................ 24
2 Materials and Methods
2.1 Bacterial and yeast strains ......................................................................... 25
2.2 Plasmids .................................................................................................... 26