High Sensitivity SELDI Analysis of NFI Interactions in Mammals, Drosophila, and Yeast
Annual rept. 15 Jul 2001-14 Jul 2002
MASSACHUSETTS GENERAL HOSPITAL BOSTON
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The role of neurofibromin in cellular growth control is complex with available data suggesting that neurofibromin is a bifunctional modulator that may regulate proliferation by coordinating the activities of numerous molecular pathways. One important step in clarifying the function of neurofibromin is the identification of critical interactors whose detection and characterization may define downstream targets of neurofibromin and regulators of its activity. The purpose of our research is identification of neurofibromin-associated proteins in multiple experimental systems using high sensitivity surface enhanced laser desorptionionization SELDI mass spectrometry. Our primary efforts have been aimed at overcoming the longstanding difficulties of manipulating normal and mutant forms of neurofibromin in mammalian cells. We have focused on using tetracycline-inducible HSV amplicon systems to re-introduce wild type neurofibromin into neurofibromin-deficient primary mouse embryo fibroblasts and human neuroblastoma cells. Our progress has been severely limited by technical difficulties that have prevented controlled exogenous neurofibromin expression in the target cells. We are now utilizing a recently available inducible amplicon system that affords much tighter regulation of transgene expression. This feature will overcome the major obstacle that has hampered our progress to date. This research will open new avenues of experimentation to clarify neurofibromins role in cellular growth control.
- Anatomy and Physiology
- Medicine and Medical Research