Worm Breeder's Gazette 15(5): 34 (February 1, 1999)
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
Department of Medical Genetics, University of British Columbia, Vancouver, B.C., V6T 1Z3
We have been characterizing the expression of ZK370.3, which shares sequence identity with human HIP1 (Huntingtin Interacting Protein) and Sla2p of yeast (Synthetic lethal with Actin Binding protein 1). ZK370.3 spans approximately 4.5 kb of genomic DNA, consisting of 10 exons and codes for a 2.7 kb message. The 923 amino acid product has a molecular weight of approximately 100 kDa, and shares identity with cytoskeletal associated protein talin.
To study the expression of C. elegans HIP1, 1000 nucleotides of the genes 5¢ upstream region, along with the first 30 nucleotides of the coding region, were fused to a GFP fusion vector (pPD95.75, kindly provided by A. Fire). The fusion construct was injected into dpy-5 animals, along with a dpy-5 rescuing clone (pCeh361, kindly provided by C. Thacker). Rescued animals were examined, and fluorescence was detected in the pharynx and tail of the animals. Expression in the pharynx appears to be limited to a set of neurons, possibly M4 and I3.
To better understand the function of the C. elegans HIP1 gene product, we conducted a yeast two-hybrid screen in collaboration with the Michael Hayden laboratory at the University of British Columbia. We used a full length C. elegans HIP1 cDNA fused to the bait vector pGBT9 (Clonetech) to screen a library kindly provided by R. Barstead. Two clones were isolated from the screen. The first shows similarity to a type II transcription factor, and the second shows similarity to the Drosophila hedgehog protein. We are currently characterizing the protein interactions.