Brant Eutzy Interactions with Nrt: A Yeast Two-Hybrid Assay Division Sciences Department Biology Year 2003 Advisor Eric Liebl

Neurotactin (Nrt) is a transmembrane protein expressed in the developing central nervous system of Drosophila embryos.  It is believed to play a role in axon pathfinding.  Yeast two-hybrid is a system that allows for the discovery of novel protein-protein interactions.  A yeast two-hybrid assay, performed on a cDNA library using the intracellular portion of Nrt as bait, revealed two hits encoding portions of the protein filamin, which is known to bundle actin in vivo.  A pulldown experiment using GST conjugated to the intracellular domain of Nrt and in vitro transcribed and translated cDNAs was performed.  Only one of the two filamin cDNAs appeared to be properly transcribed and translated and it was not observed to be pulled down by GST-Nrt.  Future experiments will likely focus on optimizing the in vitro transcription and translation and the pull down conditions along with continuing screens of the cDNA library.