Figure A4: Impaired ASH1-mRNA binding by She2p results in mislocalization of ASH1 mRNA in vivo. In situ hybridization experiments using fluorescently labeled oligonucleotides against ASH1 were performed in wild-type cells and in She2p-ΔhelixE- expressing yeast strains. a)-c) In wild-type cells ASH1 mRNA efficiently localizes to the bud tip (marked by an arrow). d)-f) In She2p-ΔhelixE-expressing Δshe2 cells no ASH1- mRNA localization to the bud tip was observed (0 % localization in n ≥ 100 cells).
(a, d) show in situ hybridization with Texas Red-labeled oligonucleotides against ASH1
mRNA, (b, e) show Hoechst nuclear staining, and (c, f) display merged signals of the respective ASH1 and nuclei stainings. This experiment was performed by Maria Schmid from the laboratory of Prof. Ralf-Peter Jansen.
Figure A5: Sedimentation profile of wild- type She2p from a sedimentation-velocity experiment. Shown is the protein distribution during a sedimentation-velocity experiment, which was performed with wild-type She2p at an intermediate concentration of 16 µM. The absorbance measured at 280 nm is plotted against the radius of the measuring cell. The curves from left to right represent the protein distribution at increasing time points. From this profile, a sedimentation coefficient of 9.64 Svedberg (S) was calculated.
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Curriculum Vitae
Marisa Müller
Personal Data
Date of birth: December 3, 1980 Place of birth: Pforzheim, Germany Citizenship: German
Marital status : Single
Education
1987 – 1991 Weiherbergschule Pforzheim – Grundschule 1991 – 2000 Reuchlin Gymnasium Pforzheim
Abitur
Academic Training
2000 – 2005 Biology studies at the Friedrich-Alexander University, Erlangen
Diploma degree in Biology
2004 – 2005 Diploma thesis at the Gene Center of the Ludwig-Maximilians- University Munich in the laboratory of Prof. Dr. Karl-Peter Hopfner “Struktur-Funktions-Untersuchungen von DNA-Reparatur
ATPasen”
2005 – 2009 Graduate studies at the Gene Center of the Ludwig-Maximilians- University Munich in the laboratory of Dr. Dierk Niessing “Characterization of She2p-dependent mRNP assembly in
Saccharomyces cerevisiae”
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Publications
Parts of this work have been published or are in process of publication:
Müller, M., Richter, K., Heuck, A., Kremmer, E., Buchner, J., Jansen, R.P., Niessing, D. (2009) Formation of She2p tetramers is required for mRNA binding, mRNP assembly, and localization
Submitted
Du, T.G., Jellbauer, S., Müller, M., Schmid, M., Niessing, D., and Jansen, R.P. (2008)
Nuclear transit of the RNA-binding protein She2 is required for translational control of localized ASH1 mRNA
EMBO Rep 9, 781-787
Other publications:
Heuck, A., Du, T.G., Jellbauer, S., Richter, K., Kruse, C., Jaklin, S., Müller, M., Buchner, J., Jansen, R.P., and Niessing, D. (2007)
Monomeric myosin V uses two binding regions for the assembly of stable translocation complexes
Proc Natl Acad Sci U S A 104, 19778-19783
Müller, M., Heuck, A., and Niessing, D. (2007)
Directional mRNA transport in eukaryotes: lessons from yeast Cell Mol Life Sci 64, 171-180
Rigali, S., Nothaft, H., Noens, E.E., Schlicht, M., Colson, S., Müller, M., Joris, B., Koerten, H.K., Hopwood, D.A., Titgemeyer, F., van Wezel, G.P. (2006)
The sugar phosphotransferase system of Streptomyces coelicolor is regulated by the GntR-family regulator DasR and links N-acetylglucosamine metabolism to the control of development Mol Microbiol 61, 1237-1251
Dürr, H., Körner, C., Müller, M., Hickmann, V., and Hopfner, K.P. (2005)
X-ray structures of the Sulfolobus solfataricus SWI2/SNF2 ATPase core and its complex with DNA
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Acknowledgements
I would like to thank my supervisor Dierk Niessing for giving me the opportunity to work in his laboratory in an excellent environment at the Gene Center. I am grateful for his guidance over the years and especially for allowing me to pursue my own strategies.
I want to thank the members of my thesis advisory committee Patrick Cramer, Klaus Förstemann, and Ralf-Peter Jansen for supporting my work over the time in a very constructive way. Especially, I am grateful to Patrick and Klaus for being the first and second examiners of my thesis.
I am grateful to Ralf-Peter Jansen for his accordance to collaborate with us on my project. He was always open for exciting discussions about the She2p mystery and I could benefit a lot from his deep knowledge on mRNA localization in yeast.
I would like to thank all members of the Niessing group for their contribution to have a really nice time in the past four years. Especially, I want to thank … Sigrun for being the “real” lab head keeping the lab running and for her great support … Alexander for being always ready for discussions on our projects, even if there was nothing to discuss ;-) … Daniela for critical comments on this thesis.
I want to thank Maria Schmid, Stephan Jellbauer, and Susanne Lange from the Jansen lab for their patience to introduce me into the work with yeast. I want to thank Klaus Richter and Johannes Buchner for performing the analytical ultracentrifugation experiments. I am very grateful to Sophia Hartung and Gregor Witte for introducing me into the SAXS technique. It was a pleasure to work with you! Alex Schele, Sophia Hartung, and Lina Karakasili I want to thank not only for having a great time together at the Gene Center, but also for nice activities outside the lab.
I am grateful to the Boehringer Ingelheim Fonds for financial support, but also for the nice atmosphere during the regular meetings and the continuous personal support. I am very glad for this fantastic experience!
Finally, and most importantly, I am deeply grateful for having a great and understanding family, who generously supported me over the last years. Thank you, Andi, for your constant support and for always being there for me!