>Would you somebody tell me why do I need to put 3-AT in yeast
>two-hybrid selection? How does exactly 3-AT work? I am a beginner in
>yeast and I need your help.
>Biological & Biochemical Sciences
>University of Houston
>Houston, TX 77204-5934
3-amino-1,2,4-triazole (3-AT) competitively inhibits imidazole glycerol
phosphate dehydratase, a His biosynthetic enzyme (Hilton et al., 1965;
Klopotowski and Wiater, 1965; Wiater et al., 1971), and therefore can limit
histidine biosynthesis and growth. For two-hybrid screens that use 3-AT, a
HIS3 reporter is employed - this gene encodes the enzyme activity inhibited
by 3AT. High expression of this reporter gene, which arises from a
successful two-hybrid interaction, can overcome the growth-inhibitory
effect of 3-AT in the medium. Therefore, the use of 3-AT and the HIS3
reporter enables positive selection for successful two-hybrid interactions.
Hilton, J. L., Kearney, P. C., and Ames, B. N. (1965). Mode of action of
the herbicide 3-amino-1,2,4-triazole (amitrole): inhibition of an enzyme
of histidine biosynthesis. Arch. Biochem. Biophys. 112, 544-547.
Klopotowski, T., and Wiater, A. (1965). Synergism of aminotriazole and
phosphate on the inhibition of yeast imidazole glycerol phosphate
dehydratase. Arch. Biochem. Biophys. 112, 562-566.
Wiater, A., Hulanicka, D., and Klopotowski, T. (1971). Structural
requirements for inhibition of yeast imidazoleglycerol phosphate
dehydratase by triazole and anion inhibitors. Acta Biochim. Pol. 18,
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