抄録
The original yeast two-hybrid system is based on the functional reconstitution of the transcriptional activator of a reporter gene by means of the interaction between bait and prey proteins, which possess DNA-binding and transactivation domains, respectively. However, in certain cases, interaction between a fusion protein with a DNA-binding domain and a bait peptide can activate the transcription of a reporter gene, irrespective of the interaction between the bait and prey proteins. Therefore, this system cannot be used to screen for prey proteins that specifically interact with the “bait” because bait peptides initiate transactivation. We used a split-Trp sensor to identify protein interactions with the C-terminal region of Arabidopsis Enhancer of Shoot Regeneration 1(ESR1), which has transactivating capabilities in yeast cells. ESR1-Interacting Candidate 1(EIC1) was identified using this system, and although its functions are unknown, EIC1 was observed to localize to nuclei when its GFP-fusion protein was expressed in onion epidermal cells. These results suggest that the split-Trp sensor could be useful in screening for proteins that interact with peptides that have transactivation domains in yeast cells.
