NR AOWY
AU Mitterreiter,S.; Gauczynski,S.; Weiss,S.
TI Life Cell Imaging of Prion Proteins and Prion Receptor Molecules
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - BR-82
PT Konferenz-Poster
AB
Life cell imaging represents a promising methodology for recording Fluorescence-labelled macromolecules such as HIV-1 (1) and prion (2) proteins in living cells. We identified the PrP interacting 37 kDa/67 kDa laminin receptor (LRP/LR) (3) as the cell surface receptor for the cellular prion protein (PrPc) (4) and HSPGs as co-factors/co-receptors for PrPc (5). Very recently, we demonstrated that LRP/LR is required for PrPsc propagation in neuronal cells (6).
In order to investigate the movement of PrPc and its receptor in mammalian cells in real time, we cloned murine LRP and PrP as a fusion with DsRed and EGFP, respectively, in recombinant Semliki-Forest-Virus vectors (pSFV-1). In addition, we constructed pmoLRP-DsRed employing pDsRed-Express-N1 (Clontech) and used pEGFP-moPrP (2), a kind gift of Dr. Kil Sun Lee, Brasil, for cell transfections.
Transfected BHK cells expressing fluorochrome-labelled prion and LRP/LR molecules were monitored by a fully automized life cell imaging microscope. The data obtained contribute to a more detailled understanding of the life cycle of prions and their receptor molecules.
1. McDonald, D., Vodicka, M. A., Lucero, G., Svitkina, T. M., Borisy, G. G., Emerman, M., and Hope, T. J. (2002) J Cell Biol 159, 441-452
2. Lee, K. S., Magalhaes, A. C., Zanata, S. M., Brentani, R. R., Martins, V. R., and Prado, M. A. (2001) J Neurochem 79, 79-87
3. Rieger, R., Edenhofer, F., Lasmézas, C. I., and Weiss, S. (1997) Nat Med 3, 1383-1388
4. Gauczynski, S., Peyrin, J. M., Haik, S., Leucht, C., Hundt, C., Rieger, R., Krasemann, S., Deslys, J. P., Dormont, D., Lasmezas, C. I., and Weiss, S. (2001) EMBO J 20, 5863-5875
5. Hundt, C., Peyrin, J. M., Haik, S., Gauczynski, S., Leucht, C., Rieger, R., Riley, M. L., Deslys, J. P., Dormont, D., Lasmezas, C. I., and Weiss, S. (2001) EMBO J 20, 5876-5886
6. Leucht, C., Simoneau, S., Rey, C., Vana, K., Rieger, R., Lasmezas, C. I., and Weiss, S. (2003) EMBO Rep 4, 290-295
AD S. Mitterreiter, S. Gauczynski, S. Weiss, Laboratorium für Molekulare Biologie - Genzentrum-Institut für Biochemie der LMU München, Germany
SP englisch
PO Deutschland