Nucleic Acids Research, Vol 24, Issue 17 3341-3347, Copyright © 1996 by Oxford University Press
CA Leanna and M Hannink
The yeast two-hybrid system is a powerful experimental approach for the
characterization of protein/ protein interactions. A unique strength of the
yeast two-hybrid system is the provision for genetic selection techniques
that enable the identification of specific protein/protein interactions. We
now report the development of a modified yeast two- hybrid system which
enables genetic selection against a specific protein/protein interaction.
This reverse two-hybrid system utilizes a yeast strain which is resistant
to cycloheximide due to the presence of a mutant cyh2 gene. This strain
also contains the wild-type CYH2 allele under the transcriptional control
of the Gal1 promoter. Expression of the wild-type Gal4 protein is
sufficient to restore growth sensitivity to cycloheximide. Growth
sensitivity towards cycloheximide is also restored by the coexpression of
the avian c-Rel protein and its I kappa B alpha counterpart, p40, as Gal4
fusion proteins. Restoration of growth sensitivity towards cycloheximide
requires the association of c- Rel and p40 at the Gal1 promoter and
correlates with the ability of the c-Rel/p40 interaction to activate
expression from the Gal1 promoter. A genetic selection scheme against
specific protein/protein interactions may be a valuable tool for the
analysis of protein/protein interactions.
ARTICLES
The reverse two-hybrid system: a genetic scheme for selection against specific protein/protein interactions
Biochemistry Department, University of Missouri-Columbia 65212, USA.
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