Dr. Ingrid Wolf

CURRICULUM VITAE

EDUCATION AND RESEARCH EXPERIENCE

since 1995: Postdoctoral fellow at the Fred Hutchinson Cancer Research Center, Seattle, USA, Lab. Dr. L.R. Rohrschneider. Project title: "Identification of protein/protein interactions involved in signal transduction pathways of the Flt3 receptor tyrosine kinase using the yeast two-hybrid system."
1990-1994: PhD-thesis at the Institute of Biochemistry, Ludwig-Maximilians- University, Munich, Germany ("summa cum laude"). Title of thesis: "The Burkitt's lymphoma receptor 1 (BLR1): Differentiation- and activation-dependent expression and identification of BLR1-binding peptides using an epitope library." Department of Prof. Dr. E.-L. Winnacker, under the supervision of Dr. M. Lipp.
1990: Diploma (Masters Degree) in Biology at the Institute of Biochemistry, Ludwig- Maximilians-University, Munich, Germany. Title of thesis: "Characterization of a Burkitt's lymphoma specific transcript." Department of Prof. Dr. E.-L. Winnacker, under the supervision of Dr. M. Lipp. 1987-1990: Study of biology at the Ludwig-Maximilians-University, Munich, Germany
1984-1987: Study of biology at the Julius-Maximilians-University, Würzburg, Germany


FELLOWSHIPS

1995-1997: Postdoctoral Fellowship from the "Deutsche Forschungsgemeinschaft"


CONFERENCE PRESENTATIONS

1991: EMBO Practical Course "Analysis and expression of neurotransmitter receptors", Organizers: P.H. Seeburg, B. Sakmann, and H. Betz, Heidelberg, Germany
1992: Posterpresentation, EMBL conference "Oncogenes and growth control", Heidelberg, Germany
1993: Oral presentation, 7. AEK-Symposium of the German Cancer Society, Heidelberg, Germany
1993: Posterpresentation, Keystone Symposia "B and T cell lymphomas", Copper Mountain, Colorado, USA
May, 1997: Posterpresentation, Congress of Molecular Medicine, Berlin, Germany
June, 1997: Oral presentation, Thirteenth Annual Meeting on Oncogenes - Oncogenes, Growth, and Development, Frederick, MD, USA


TECHNICAL SKILLS

• all general methods in molecular biology
• screening cDNA and genomic libraries
• work with RNA and mRNA
• yeast two-hybrid system
• in vitro selection methods (epitope library, DNA binding site selection)
• immunoprecipitation, western blotting, immunofluorescence
• protein expression in various systems
• tissue culture
• generation of antibodies
• various reporter assay systems (CAT, luciferase, beta-galactosidase)

PUBLICATIONS

Ilangumaran, S., Wolf, I., Gertler, F., LaRose, J., Bhoi, P., Paganin, C., Anafi, M., Rohrschneider, L., and Rottapel, R. (1999) Functional interaction between the c-Abl tyrosine kinase and the B-cell lineage receptor Flt3/Flk2. submitted.

Wolf, I. and Rohrschneider, L. (1999) Fiz1, a novel zinc finger protein interacting with the receptor tyrosine kinase Flt3. J. Biol. Chem., 274: 21478-21484.

Pevzner, V., Wolf, I., Burgstahler R., Förster, R., and Lipp. M. (1999) Regulation of expression of chemokine receptor BLR1/CXCR5 during B cell maturation. Curr. Top. Microbiol. Immunol., 246: 79-84.

Wolf, I., Pevzner, V., Kaiser, E., Bernhardt, G., Claudio, E., Siebenlist, U., Förster, R., and Lipp. M. (1998) Downstream activation of a TATA-less promoter by Oct-2, Bob1, and NF-kB directs expression of the homing receptor BLR1 to mature B cells. J. Biol. Chem., 273: 28831-28836.

Förster, R., Kaiser, E., Wolf, I., and Lipp, M. (1994) Selective expression of the murine homologue of the G-protein-coupled receptor BLR1 in B cell differentiation, B cell neoplasia, and defined areas of the cerebellum. Cell. Mol. Biol., 40: 381-387.

Kaiser, E., Förster, R., Wolf, I., Ebensperger, C., Kuehl, M., and Lipp, M. (1993) The G protein-coupled receptor BLR1 is involved in murine B cell differentiation and is also expressed in neuronal tissues. Eur. J. Immunol., 23: 2532-2539.

Dobner, T., Wolf, I., Emrich, T., and Lipp, M. (1992) Differentiation-specific expression of a novel G protein-coupled receptor from Burkitt's lymphoma. Eur. J Immunol., 22: 2795-2799.

Dobner, T., Wolf, I., Mai, B., and Lipp, M. (1991): A Novel Divergently Transcribed Human Histone H2A/H2B Gene Pair. DNA Sequence, 1: 409-413.

RESEARCH INTERESTS

Within the hematopoietic system, a pluripotent stem cell gives rise to eight different hematopoietic lineages. Cytokines, growth factors and their receptors, respectively, play an important role in this process. The hematopoietic class III receptor tyrosine kinases (RTK) Kit, Fms, and Flt3 and their ligands SCF, M-CSF, and FL are involved at different stages of this process and are important for the proliferation and differentiation of different lineage precursors. My research focusses on the Flt3 RTK which is structurally related to the Fms and Kit RTKs. All three receptors use well characterized molecules in their signalling pathway such as the p85 subunit of PI-3'-kinase, PLCgamma, Shc, Grb2 etc. However, to understand the different functions of these RTKs it is necessary to learn more about substrates used specifically by one RTK but not the others. Using the Flt3 cytoplasmic domain as bait in a two hybrid screen I isolated several SH2-domain containing signalling molecules such as p85, Shc, Nck, and 3BP2. Binding of a SH2-domain to phosphotyrosine residues of the activated receptor is a well characterized interaction. In addition, I identified the novel protein Fiz1 (Flt3 interacting zinc finger protein 1) containing 11 zinc finger domains. This interaction is not dependent on binding of a SH2-domain to a phosphotyrosine residue and reveals a new interaction mechanism of a RTK and a signalling molecule. Binding of Fiz1 to Flt3 is not dependent on the kinase activity of Flt3 and I confirmed the interaction using different in vitro assays. Further studies have shown, that Fiz1 binds to the catalytic kinase domain of Flt3 and not to any of the related RTKs such as Fms and Kit. Anti-Fiz1 antibodies detect a single 55-60 kDa protein which is localized in the nucleus as well as in the cytoplasm. Immunofluorescence studies further have shown that Fiz1 partially colocalizes with the transferrin receptor, a marker for early endosomes. Therefore, early endosomes might be the site where Fiz1 does interact with Flt3. It will be interesting to analyze whether Fiz1 plays a role in receptor degradation or recycling to the cell surface.