Exploring Neurofibromin Function in a Yeast Model of NF1
Abstract
We hypothesized that the NF1 protein Neurofibromin has functions in addition to its Ras-GAP activity and proposed to develop a yeast NF1 model to define new functions as well as novel regulators of known functions. In year 1 of this project, we categorized genetic hits that we recovered from yeast screens. We divided hits into Ras-dependent and Ras-independent categories. To do this, we used two approaches. First, we assayed mutant yeast strains from our screens by idodine staining, providing an indirect readout for ras signaling levels. For strains that showed elevated ras signaling, we next used Ras-GTP pull-down assays to define if the genes functioned upstream or downstream of ras. The most significant finding during the first year of this project is that we defined which of our genes that interact with IRA1 or IRA2 are involved in ras signaling and which are not. Furthermore, of the genes involved in ras signaling, we now have evidence placing these either upstream or downstream of ras. These data will facilitate future years of funding for this project, as we define if the genetic modifiers interact physically with Ira1 and/or Ira2, as well as generating transgenic flies to validate our results in Drosophila.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2009
- Accession Number
- ADA519204
Entities
People
- Aaron Gitler
Organizations
- University of Pennsylvania