Gal80 Dimerization and the Yeast GAL Gene Switch

Pilauri, Vepkhia; Bewley, Maria C.; Diep, Cuong Q.; Hopper, James E.

doi:10.1534/genetics.104.036723

Cited by 60 publications

(70 citation statements)

References 80 publications

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“…A comparison of the ␣-carbon traces for Gal80p and Z. mobilis oxidoreductase (Fig. 2a) demonstrates that these two proteins superimpose with a rootmean-square deviation of 2.0 Å for 307 structurally equivalent ␣-carbons, which is remarkable given their limited amino acid sequence identity of ϳ13% (16). It has been postulated that Lys-129 and Tyr-217 are involved in the catalytic mechanism of the oxidoreductase (13).…”

Section: Resultsmentioning

confidence: 93%

“…Several recent and insightful mutational analyses of ScGal80p have been performed (16,18). In one of these studies, variants of the protein from S. cerevisiae were uncovered that were defective only in Gal4p-or in Gal3p binding (16).…”

Section: Resultsmentioning

confidence: 99%

“…In one of these studies, variants of the protein from S. cerevisiae were uncovered that were defective only in Gal4p-or in Gal3p binding (16). Given the amino acid sequence homology between the S. cerevisiae and K. lactis proteins, we have mapped these mutations onto the KlGal80p structure (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The mutations at Gly-153 and Gly-184, however, are particularly interesting because they are separated by ϳ17 Å and lie on either side of a large cleft formed by the COOHterminal end of the ␤-sheet in the Rossmann fold and an ␣-helix defined by Ser-211 to Ile-222. Two additional mutations have been identified in ScGal80p that result in defective Gal4p-binding: A309T and G310D (16). The corresponding amino acids in FIGURE 2.…”

Section: Resultsmentioning

confidence: 99%

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Understanding a Transcriptional Paradigm at the Molecular Level

Thoden

Sellick

Reece

et al. 2007

Journal of Biological Chemistry

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In yeast, the GAL genes encode the enzymes required for normal galactose metabolism. Regulation of these genes in response to the organism being challenged with galactose has served as a paradigm for eukaryotic transcriptional control over the last 50 years. Three proteins, the activator Gal4p, the repressor Gal80p, and the ligand sensor Gal3p, control the switch between inert and active gene expression. Gal80p, the focus of this investigation, plays a pivotal role both in terms of repressing the activity of Gal4p and allowing the GAL switch to respond to galactose. Here we present the three-dimensional structure of Gal80p from Kluyveromyces lactis and show that it is structurally homologous to glucose-fructose oxidoreductase, an enzyme in the sorbitol-gluconate pathway. Our results clearly define the overall tertiary and quaternary structure of Gal80p and suggest that Gal4p and Gal3p bind to Gal80p at distinct but overlapping sites. In addition to providing a molecular basis for previous biochemical and genetic studies, our structure demonstrates that much of the enzymatic scaffold of the oxidoreductase has been maintained in Gal80p, but it is utilized in a very different manner to facilitate transcriptional regulation.

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Section: Resultsmentioning

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Understanding a Transcriptional Paradigm at the Molecular Level

Thoden

Sellick

Reece

et al. 2007

Journal of Biological Chemistry

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“…ScPD2 was derived from ScVP2-103 (Pilauri et al 2005) by disruption of the GAL1 gene. Sc750 and Sc751 were derived from Sc724 (Blank et al 1997) by disruption of the GAL80 gene and integration of the GAL80 S-0 and GAL80 S-1 alleles, respectively.…”

Section: Methodsmentioning

confidence: 99%

Genetic Evidence for Sites of Interaction Between the Gal3 and Gal80 Proteins of the Saccharomyces cerevisiae GAL Gene Switch

Diep¹,

Tao²,

Pilauri³

et al. 2008

Genetics

Self Cite

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Galactose-activated transcription of the Saccharomyces cerevisiae GAL genes occurs when Gal3 binds the Gal4 inhibitor, Gal80. Noninteracting variants of Gal3 or Gal80 render the GAL genes noninducible. To identify the binding determinants for Gal3's interaction with Gal80 we carried out GAL3-GAL80 intergenic suppression analyses and selected for new GAL3 mutations that impair the Gal3-Gal80 interaction. We show that a GAL3 C -D368V mutation can suppress the noninducibility due to a GAL80 S-1 -G323R mutation, and a GAL80-M350C mutation can suppress the noninducibility due to a gal3-D111C mutation. A reverse two-hybrid selection for GAL3 mutations that impair the Gal3-Gal80 interaction yielded 12 singleamino-acid substitutions at residues that are predicted to be surface exposed on Gal3. The majority of the affected Gal3 residues localized to a composite surface that includes D111 and a sequence motif containing D368, which has been implicated in interaction with Gal80. The striking colocalization of intergenic suppressor residues and Gal80 nonbinder residues identifies a Gal3 surface that likely interacts with Gal80.

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Ectopic expression in commonly used transgenic DrosophilaGAL4 driver lines

Winant,

Buhler,

Callaerts

2024

Genesis

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SummaryTransgenic tools such as the GAL4/UAS system in Drosophila have been used extensively to induce spatiotemporally controlled changes in gene expression and tissue‐specific expression of a range of transgenes. We previously discovered unexpected expression of the commonly used dilp2‐GAL4 line in tracheal tissue which significantly impacted growth phenotypes. We realized that few GAL4 lines have been thoroughly characterized, particularly when considering transient activity that may have significant impact on phenotypic readouts. Here, we characterized a further subset of 12 reportedly tissue‐specific GAL4 lines commonly used in genetic studies of development, growth, endocrine regulation, and metabolism. Ten out of 12 GAL4 lines exhibited ectopic activity in other larval tissues, with seven being active in the larval trachea. Since this ectopic activity may result in phenotypes that do not depend on the manipulation in the intended target tissue, it is recommended to carefully analyze the outcome while taking this aspect into consideration.

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Gal80 Dimerization and the Yeast GAL Gene Switch

Abstract: The

Cited by 60 publications

References 80 publications

Understanding a Transcriptional Paradigm at the Molecular Level

Understanding a Transcriptional Paradigm at the Molecular Level

Genetic Evidence for Sites of Interaction Between the Gal3 and Gal80 Proteins of the Saccharomyces cerevisiae GAL Gene Switch

Ectopic expression in commonly used transgenic DrosophilaGAL4 driver lines

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