148 The work presented in this thesis has been a joint endeavor of the laboratories of Prof. Dirk

Görlich (MPI for Biophysical Chemistry, Göttingen), Prof. Ralf Ficner (Georg-August-Universität Göttingen), Prof. Michael Sattler (Technische (Georg-August-Universität und Helmholtz Zentrum München), Dr. Raymond Poot (Erasmus Medical Center, Rotterdam, Netherlands), Dr.

Robbert Rottier (Erasmus Medical Center, Rotterdam, Netherlands) and Prof. Giulio Superti-Furga (CeMM, Vienna, Austria). All contributors to the presented studies are listed on the chapters' title pages or in the acknowledgment sections. For the sake of brevity, the following statement only summarizes the contributions by the thesis' author.

CHAPTER 2

Gontan, C., Güttler, T., Engelen, E., Demmers, J., Fornerod, M., Grosveld, F. G., Tibboel, D., Görlich, D., Poot, R. A. & Rottier, R. J. (2009) Exportin 4 mediates a novel nuclear import pathway for Sox family transcription factors. J Cell Biol 185, 27-34.

C. Gontan and T. Güttler contributed equally to this work.

T. Güttler conceived central nuclear import experiments and prepared reagents (including the α-X. laevis Exp4 antibody, many fluorescent probes and transport receptors). He performed part of the binding and import assays. T. Güttler contributed to the interpretation of the data, wrote the first draft of the manuscript and helped to complete the paper for publication (including the preparation of figures).

CHAPTER 3

Monecke, T., Güttler, T., Neumann, P., Dickmanns, A., Görlich, D. & Ficner, R. (2009) Crystal structure of the nuclear export receptor CRM1 in complex with Snurportin1 and RanGTP. Science 324, 1087-1091.

T. Monecke and T. Güttler contributed equally to this work.

T. Güttler designed part of the biochemical experiments. He prepared all proteins (with the exception of Snurportin1 used for the crystallization experiments), some with the help of D.

Deichsel. T. Güttler established constructs and conditions for the assembly of stable, chromatographically homogeneous and crystallizable export complexes. T. Güttler contributed to screening for crystallization conditions. He performed all biochemical assays described in the main body and in the supplements of the paper. He contributed to the interpretation of the data and to writing of the manuscript (including the preparation of figures).

149 Sattler, M., Görlich, D. (2010) NES consensus redefined by structures of PKI-type and Rev-type nuclear export signals bound to CRM1. Nat Struct Mol Biol, accepted for publication (on condition of editorial changes in the text).

T. Güttler, T. Madl and P. Neumann contributed equally to this work.

Note added for publication of this thesis:

The citation of the article's print version is Nat Struct Mol Biol 17, 1367-1376.

T. Güttler conceived part of this study. He designed expression constructs and optimized the recombinant protein expression and purification. All proteins have been prepared by T.

Güttler with the help of D. Deichsel. T. Güttler carried out all binding and export experiments, established the deuteration conditions for CRM1, performed isotope labeling and prepared all samples for the NMR-spectroscopic experiments. T. Güttler reconstituted and crystallized all export complexes and contributed to data collection and structure solving.

T. Güttler prepared all figures shown in the manuscript (Fig. 3 and Supplementary Fig. 4+5 together with T. Madl). T. Güttler contributed to data interpretation and to writing of the manuscript.

Hantschel, O., Wiesner, S., Güttler, T., Mackereth, C. D., Rix, L. L., Mikes, Z., Dehne, J., Görlich, D., Sattler, M. & Superti-Furga, G. (2005) Structural basis for the cytoskeletal association of Bcr-Abl/c-Abl. Mol Cell 19, 461-473.

T. Güttler conceived part of the nuclear transport experiments. T. Güttler prepared all reagents for the nuclear transport part of the paper and performed all pulldown and export assays (Fig.

3). To verify the data, he carried out numerous experiments not shown in the paper, including microinjection assays with X. laevis oocytes and immunofluorescence experiments. T. Güttler contributed to interpretation of the data and to writing of the paper.

Göttingen, August 2010

Thomas Güttler Prof. Dr. Dirk Görlich

150

Name Thomas Güttler

Day and place of birth July 12, 1978; Löbau (Germany)

Nationality German

Education

2007-2010 Max Planck Institute for Biophysical Chemistry, Göttingen (Germany) and Georg-August-Universität Göttingen

GGNB Program "Biomolecules: Structure-Function-Dynamics"

Doctoral studies in the laboratory of Prof. Dirk Görlich

2004-2007 Ruprecht-Karls-Universität Heidelberg, ZMBH (Germany) Doctoral studies in the laboratory of Prof. Dirk Görlich

2001-2003 Ruprecht-Karls-Universität Heidelberg (Germany)

Diploma studies in the International Master's Program "Molecular and Cellular Biology"

Diploma/Master Thesis in the laboratory of Prof. Dirk Görlich (ZMBH)

"Nuclear export of translation factors as a general mechanism to confine translation to the cytoplasm"

2001 University of Copenhagen (Denmark)

Workshops on biomembranes, membrane proteins and signal transduction 2000-2001 University of Mississippi (Oxford/MS, U.S.A.)

Studies of biochemistry and microbiology

Student research project in the laboratory of Dr. Lidija Halda-Alija

"Quorum sensing by wetland rhizobacteria"

1998-2000 Friedrich-Schiller-Universität Jena (Germany) Undergraduate studies in biology

Pre-degree ("Vordiplom") in biology

1997 Geschwister-Scholl-Gymnasium Löbau (Germany) Abitur

151 Güttler, T. *, Madl, T. *, Neumann, P. *, Deichsel, D., Corsini, L., Monecke, T., Ficner, R., Sattler, M., Görlich, D. (2010). NES consensus redefined by structures of PKI-type and Rev-type nuclear export signals bound to CRM1. Nat. Struct. Mol. Biol. accepted for publication (on condition of editorial changes in the text).

Note added for publication of this thesis:

The citation of the article's print version is Nat Struct Mol Biol 17, 1367-1376.

Mohr, D., Frey, S., Fischer, T., Güttler, T., and Görlich, D. (2009). Characterisation of the passive permeability barrier of nuclear pore complexes. EMBO J. 28, 2541-53.

Monecke, T.*, Güttler, T.*, Neumann, P., Dickmanns, A., Görlich, D., and Ficner, R. (2009).

Crystal structure of the nuclear export receptor CRM1 in complex with Snurportin1 and RanGTP. Science 324, 1087-1091.

Gontan, C.*, Güttler, T.*, Engelen, E., Demmers, J., Fornerod, M., Grosveld, F., Tibboel, D., Görlich, D., Poot, R.A., and Rottier, R. (2009). Exportin 4 mediates a novel nuclear import pathway for Sox family transcription factors. J. Cell Biol., 185, 27-34.

Hantschel, O., Wiesner, S., Güttler, T., Mackereth, C. D., Rix, L. L., Mikes, Z., Dehne, J., Görlich, D., Sattler, M., and Superti-Furga, G. (2005). Structural basis for the cytoskeletal association of Bcr-Abl/c-Abl. Mol. Cell 19, 461-73.

Bono, F., Ebert, J., Unterholzner, L., Güttler, T., Izaurralde, E., and Conti, E. (2004).

Molecular insights into the interaction of PYM with the Mago-Y14 core of the exon junction complex. EMBO Rep. 5, 304-10.

Kruse, C., Jaedicke, A., Beaudouin, J., Böhl, F., Ferring, D., Güttler, T., Ellenberg, J., and Jansen, R. P. (2002). Ribonucleoprotein-dependent localization of the yeast class V myosin Myo4p. J. Cell Biol. 159, 971-82.