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doi:10.1534/genetics.105.050807
A more recent version of this article appeared on January 1, 2006.
REGULAR RESEARCH PAPERS |
Intragenic suppression of Gal3C interaction with Gal80 in the Saccharomyces cerevisiae GAL gene switch
Cuong Q. Diep 1, Gang Peng 2, Maria Bewley 1, Vepkhia Pilauri 1, Ira Ropson 1 and James E Hopper 1*
1 Pennsylvania State University, College of Medicine
2 University of Oregon, Institute of Neuroscience
* To whom correspondence should be addressed. E-mail: jhopper{at}psu.edu.
Submitted on September 7, 2005
Revised on October 4, 2005
Accepted on 5 October 2005
Gal4-mediated activation of GAL gene transcription in Saccharomyces cerevisiae requires the interaction of Gal3 with Gal80, the Gal4 inhibitor protein. While it is known that galactose and ATP activates Gal3 interaction with Gal80, neither the mechanism of activation nor the surface that binds to Gal80 is known. We addressed this through intragenic suppression of GAL3C alleles that cause galactose-independent Gal3-Gal80 interaction. We created a new allele, GAL3SOC, and showed that it suppressed a new GAL3C allele. We tested the affect of GAL3SOC on several newly isolated and existing GAL3C alleles that map throughout the gene. All except one GAL3C allele, D368V, were suppressible by GAL3SOC. GAL3SOC and all GAL3C alleles were localized on a Gal3 homology model that is based on the structure of the highly related Gal1 protein. These results provide evidence for allosterism in the galactose- and ATP-activation of Gal3 binding to Gal80. In addition, because D368V and residues corresponding to Gal80-nonbinder mutations co-localized to a domain that is absent in homologous proteins that do not bind to Gal80, we suggest that D368 is a part of the Gal80-binding surface.
Key Words: GAL gene switch, Gal3, Gal80, Intragenic suppression
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  C. Q. Diep, X. Tao, V. Pilauri, M. Losiewicz, T. E. Blank, and J. E. Hopper Genetic Evidence for Sites of Interaction Between the Gal3 and Gal80 Proteins of the Saccharomyces cerevisiae GAL Gene Switch Genetics, February 1, 2008; 178(2): 725 - 736. [Abstract] [Full Text] [PDF]
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