Characterization of minimalist fusion hybrids of Arnt in a yeast one-hybrid system.
Contributors: University of Toronto.  Dept. of Chemistry.
Published: 2007
Language: eng
Number of pages: 90
ISBN-13: 9780494274378
Description: Minimalist hybrids comprising the DNA-binding domain of bHLH/PAS protein Arnt and leucine zipper of bZIP protein C/EBP were designed to bind E-box (5'CACGTG). Swapping of the Arnt PAS domain for C/EBP leucine zipper generates the hybrid ArntbHLH-C/EBP, which binds to E-box modestly in the yeast one-hybrid system. Positive binding was also observed when ArntbHLH is fused to the Max leucine zipper; therefore, leucine zippers from different protein families can replace the Arnt PAS domain for effecting homodimerization. Removal of the Arnt HLH domain or the nonnative leucine zipper yields nonfunctional proteins, suggesting that two dimerization domains work in conjunction of each other and are essential for DNA-binding. Additionally, random mutants of our fusion hybrids of Arnt were screened for improved E-box binding function in a modified yeast one-hybrid system. Our minimalist design strategy successfully reduced the size and structural complexity of native Arnt bHLH/PAS to a functional bHLH/ZIP-like structure.
Notes: Source: Masters Abstracts International, Volume: 45-06, page: 3059.

Thesis (M.Sc.)--University of Toronto, 2007.

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