Research Group Signal Transduction

Projects in brief

Molecular and biochemical characterization of LMP1 signal transduction pathways and their role in the control of B-cell proliferation, survival and transformation. Currently, we focus on the NF-kappaB, JNK and PI3-kinase pathways.

The role of secondary protein modifications (phosphorylation, ubiquitinylation, SUMOylation)

The physiological role of critical LMP1 signaling intermediates in cellular signaling and function of TNF-receptors, CD40 and Toll-like receptors. Differences in viral transforming signaling versus physiological signaling.

Biochemical purification and characterization of the LMP1 signaling complex by functional proteomics. Functional characterization of newly identified interaction partners.

The role of the germinal center kinase TNIK in LMP1 and lymphocyte signaling. TNIK's functions in hematopoiesis and immune response using TNIK knockout-mice.

Translational research: Development of strategies to interfere with EBV-induced tumors based on the specific inhibition of LMP1-induced signal transduction. We develop high-throughput screening protein-protein interaction assays to identify small molecule inhibitors of LMP1's interaction with its critical signaling intermediates.

Techniques established in the laboratory

All standard molecular biology methods

Molecular cloning, PCR, qPCR, DNA-, RNA-purification, Southern Blot etc.

Protein biochemistry and proteomics

  • Immunoblotting
  • Immunoprecipitation methods
  • Membrane lipid raft preparation and analysis
  • ELISA
  • Analysis of protein complexes in vitro and in vivo
  • In vitro kinase assays
  • Analysis of ubiquitinylation and SUMOylation
  • In vitro transcription / translation of proteins
  • Expression and purification of recombinant proteins in E.coli
  • Biacore protein-protein interaction studies
  • Peptide array interaction studies
  • Development of high-throughput screening assays for protein-protein interations
  • Proteomics (sample preperation, protein complexes and secondary modifications)

Cell culture and in vivo methods

  • Transfection of mammalian cells (electroporation, lipofection)
  • Virus preparation and infection (Epstein-Barr virus and retroviral vectors)
  • Immunofluorescence, confocal microscopy
  • In vivo fluorescence methods (digital live cell imaging)
  • Flow cytometry
  • Cell cycle and apoptosis
  • Gene knockout in human somatic cells by homologous recombination in cell culture
  • RNA interference (siRNA and RNAi vectors)
  • Transgenic mouse technology
  • Mouse models
  • Analysis of the hematopoietic system of mice

Our work is supported by grants from the Deutsche Forschungsgemeinschaft (DFG) and the Helmholtz-Gemeinschaft Deutscher Forschungszentren (HGF).

Our project involving the characterization of the germinal center kinase TNIK in hematopoiesis and signaling is part of the collaborative research center 684 (SFB 684) "Molecular Mechanisms of Normal and Malignant Hematopoiesis".