Interaction of GAL4 and GAL80 gene regulatory proteins in vitro.

Lue, Neal F.; Chasman, Daniel I.; Buchman, A R; Kornberg, Roger D.

doi:10.1128/mcb.7.10.3446

Cited by 163 publications

(92 citation statements)

References 37 publications

(35 reference statements)

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“…The resulting association and dissociation rate constants affecting all proteinprotein interactions, and also DNA -protein, are provided in Table 3 (Equations (2), (3), (4) (5) and (7)). The dissociation constant K D for the complex G4dG80d is equivalent to 0.85x10 -9 M, close to the 0.3x10 -9 M reported by Melcher & Xu [43], or the 5x10 -9 M reported by Lue et al [76]. However, the affinity of G80 proteins to produce a dimer is much lower (7.9x10 -8 M) than the reported by Melcher & Xu [43] (1-3x10 -10 M).…”

Section: Direct or Indirect Induction?contrasting

confidence: 41%

Evolution of design principles in biochemical networks

et al. 2004

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Computer modelling and simulation are commonly used to analyse engineered systems.Biological systems differ in that they often can not be accurately characterized, so simulations are far from exact. Nonetheless , we argue in this paper that evolution results in recurring, dynamic organizational principles in biological systems, and that simulation can help to identify them and analyse their dynamic properties. As a specific example, we present a dynamic model of the galactose utilization pathway in yeast, and highlight several features of the model that embody such "design principles".

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Section: Direct or Indirect Induction?contrasting

confidence: 41%

Evolution of design principles in biochemical networks

et al. 2004

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“…For example, the herpes viral protein VP16 augments transcription of particular genes by complexing with specific DNA-binding proteins and providing an acidic activation domain (Gerster and Roeder 1988;Preston et al 1988;Triezenberg et al 1988a,b). Mechanisms by which activators respond to regulatory signals are diverse and include sites in the protein that bind effectors (e.g., glucocorticoid receptor; Rasconi and Yamamoto 1987), sites in the protein that are modified covalently (e.g, ADRl; Cherry et al 1988), regulation in the synthesis of the activator (e.g., GCN4; Hiimebusch 1984; Thireos et al 1984), and regulation by direct interaction with another protein (e.g., GAL4-GAL80; Lue et al 1987). Any component in the regula tory complex at the DNA may be targeted for regulation by these means.…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of HAP4: a third component of the CCAAT-bound HAP2/HAP3 heteromer.

Forsburg¹,

Guarente²

1989

Genes Dev.

413

348

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The CYCl gene of Saccharomyces cerevisiae is positively regulated by the HAP2 and HAPS proteins, which form a heteromeric complex that binds to a CCAAT box in the upstream activation site, UAS2, and which activate transcription in a nonfermentable carbon source. We carried out a genetic analysis to identify additional trans-acting regulatory factors exerting their effects through UAS2. We present the identification and characterization of a new locus, HAP4, which is shown to encode a subunit of the DNA-binding complex at UAS2. In the bap4 mutant, the binding of HAP2 and HAP3 (HAP2/3) is not observed in vitro. The HAP4 gene is regulated transcriptionally by a carbon source, suggesting that it encodes a regulatory subunit of the bound complex. The sequence of HAP4 shows a highly acidic region, which innactivated the protein when deleted. Replacement of this region with the activation domain of GAL4 restored activity, suggesting that it provides the principal activation domain to the bound HAP2/3/4 complex.

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“…2 and d), and the CDC7-overproducing strains SEY6210/pG12/pSYC756 (lanes b and e), and SEY6210/pG12/pSYC758 (lanes c and f) grown in synthetic complete (SC) medium (minus uracil and leucine) plus 2% glucose were diluted 1:10 in SC medium (minus uracil and leucine) containing either 2% glucose (lanes a-c) or 2% galactose (lanes d-f). Plasmid pG12 provides the GAL4 protein essential for expression from the GAL10 promoter (18). After 8 hr at 30°C, whole yeast lysates were prepared by disruption of the cells with glass beads and clarified by centrifugation.…”

Section: Methodsmentioning

confidence: 99%

The CDC7 protein of Saccharomyces cerevisiae is a phosphoprotein that contains protein kinase activity.

Yoon

Campbell

1991

Proc. Natl. Acad. Sci. U.S.A.

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The CDC7 protein of Saccharomyces cerevisiae may be involved in the G1/S-phase transition and/or in the initiation of mitotic DNA synthesis. The CDC7 gene has two in-frame AUG codons as possible translation start sites, which would produce 58-and 56-kDa proteins, respectively. Both p58 and p56 derived from recombinant plasmids complement the temperature-sensitive growth defect of the cdc7-1 allele. To determine the biochemical function of the CDC7 protein, the CDC7 gene was cloned and polyclonal antibodies were produced against the CDC7 protein. CDC7 immune complexes prepared from yeast with these antibodies phosphorylate histone H1. Kinase activity is thermolabile in strains carrying the cdc7-1 temperature-sensitive mutant allele and is elevated >10-fold in strains carrying plasmids overexpressing either p56 or p58, confirming that the kinase in the immunoprecipitates is the CDC7 gene product. In addition, we show that CDC7 is a phosphoprotein itself. Indirect immunofluorescence and biochemical fractionation show that the CDC7 protein is present at relatively high concentrations in the nucleus compared with the cytoplasm, suggesting that nuclear proteins may be substrates for the CDC7 protein.Two major events define the eukaryotic cell cycle: replication of the chromosomes during S phase and segregation of the chromosomes during mitosis. Replication and segregation are separated by two gap periods, G1 and G2, during which the cells prepare for these events. There are also two major control points for the cell cycle in G1 and G2, the point of commitment to DNA synthesis in G1 and the regulation of progression into mitosis in G2. Although we have made great strides in understanding mitotic control in the past three years, relatively little is known about the events in G1 that lead to S phase. In yeast, the point of commitment to DNA synthesis is defined as "Start", early in G1. DNA synthesis does not ensue immediately after Start, however, and it is not clear whether the lag is due to assembly of the replication apparatus or to a cascade of controls initiated at Start. CDC28, CDC4, and CDC7 define a dependent series of events set in motion at Start (1-3). CDC28 encodes a protein kinase subunit, the analog of the highly conserved cdc2+/ MPF kinase subunit involved in regulation of mitosis (3).CDC4 is homologous to one subunit of the signal transducing protein, transducin (4). The DNA sequence of the CDC7 gene predicts a protein that has homology to catalytic domains of the protein kinases CDC2/CDC28, NIM1, and a number of kinase-related transforming proteins (5). The CDC7 sequence differs from that of other protein kinases in the so-called phosphate acceptor domain, however (5, 6). We have been interested in the role of CDC7 in the events linking Start and the initiation of DNA replication. Mutations in CDC7 appear to block a precondition for DNA synthesis because cells carrying these lesions cannot start new rounds of DNA replication after a shift from permissive to nonpermissive temperature (7,8). The cd...

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Interaction of GAL4 and GAL80 gene regulatory proteins in vitro.

Cited by 163 publications

References 37 publications

Evolution of design principles in biochemical networks

Evolution of design principles in biochemical networks

Identification and characterization of HAP4: a third component of the CCAAT-bound HAP2/HAP3 heteromer.

The CDC7 protein of Saccharomyces cerevisiae is a phosphoprotein that contains protein kinase activity.

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