Open AccessInteraction of GAL4 and GAL80 gene regulatory proteins in vitro.
Author(s) -
Neal F. Lue,
Daniel I. Chasman,
A R Buchman,
Roger D. Kornberg
Publication year - 1987
Publication title -
molecular and cellular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.14
H-Index - 327
eISSN - 1067-8824
pISSN - 0270-7306
DOI - 10.1128/mcb.7.10.3446
Subject(s) - biology , fusion protein , immunoprecipitation , upstream activating sequence , protein a/g , ura3 , microbiology and biotechnology , dna binding protein , biochemistry , vesicle associated membrane protein 8 , gene product , dna , hspa2 , peptide sequence , antiserum , binding protein , saccharomyces cerevisiae , plasmid , gene , transcription factor , membrane protein , gene expression , recombinant dna , genetics , antigen , promoter , membrane
The GAL80 protein of Saccharomyces cerevisiae, synthesized in vitro, bound tightly to GAL4 protein and to a GAL4 protein-upstream activation sequence DNA complex, as shown by (i) coimmunoprecipitation of GAL4 and GAL80 proteins with anti-GAL4 antiserum, (ii) an electrophoretic mobility shift of a GAL4 protein-upstream activation sequence DNA complex upon the addition of GAL80 protein, and (iii) GAL4-dependent binding of GAL80 protein to upstream activation sequence DNA immobilized on Sepharose beads. Anti-GAL4 antisera were raised against a GAL4-URA3 fusion protein, which could be purified to homogeneity in a single step with the use of an affinity chromatographic procedure for the URA3 gene product.