3-AT

Matthew Sachs msachs at ADMIN.OGI.EDU
Tue Jan 21 13:21:50 EST 1997


>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.
>
>Karen Yoas
>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,
289-297.





-----------------------------------------------------------
Matthew Sachs
Department of Chemistry, Biochemistry and Molecular Biology
Oregon Graduate Institute of Science and Technology
20000 NW Walker Road
P.O. Box 91000
Portland, OR  97291-1000
503 690-1487 Phone
503 690-1464 Fax
msachs at admin.ogi.edu





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