Green fluorescent protein inSaccharomyces cerevisiae: Real-time studies of theGAL1 promoter

Li, Jincai; Wang, Shu; VanDusen, William J.; Schultz, Loren D.; George, Hugh A.; Herber, Wayne K.; Chae, Hee Jeong; Bentley, William E.; Rao, Govind

doi:10.1002/1097-0290(20001020)70:2<187::aid-bit8>3.0.co;2-h

Cited by 77 publications

(47 citation statements)

References 37 publications

(38 reference statements)

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“…Li et al [21] showed that this graded induction has a plateau reached at galactose concentrations between 1 g/L and 3 g/L (5.5 and 16.7 mM respectively), and maintained up to 20 g/L of applied external galactose. They also showed that at 0.1 g/L (0.55 mM) of external galactose, GAL gene expression is only 10% of the maxi mum.…”

Section: The Galactose Utilization Modelmentioning

confidence: 97%

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: The Galactose Utilization Modelmentioning

confidence: 97%

Evolution of design principles in biochemical networks

et al. 2004

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“…We employ a reporter green fluorescence protein (gfp) expressed from a GAL promoter and high-resolution fluorescence microscopy to quantify the protein distribution in the population, and to characterize both their steady states and their dynamic responses to various perturbations. Previous studies have shown that this method accurately reflects the pattern of the native GAL system proteins [12,35]. Direct analysis of microscope images allows us to overcome artifacts that may occur while using automated analysis methods [36] and to analyze in detail the physiological properties of the population (see below).…”

Section: Introductionmentioning

confidence: 99%

Dynamics of protein distributions in cell populations

2006

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A population of cells exhibits wide phenotypic variation even if it is genetically homogeneous. In particular, individual cells differ from one another in the amount of protein they express under a given regulatory system under fixed conditions. Here we study how protein distributions in a population of the yeast S. cerevisiae are shaped by a balance of processes: protein production-an intracellular process-and protein dilution due to cell division-a population process. We measure protein distributions by employing reporter green fluorescence protein (gfp) under the regulation of the yeast GAL system under conditions where it is metabolically essential. Cell populations are grown in chemostats, thus allowing control of the environment and stable measurements of distribution dynamics over many generations. Despite the essential functional role of the GAL system in a pure galactose medium, steady-state distributions are found to be universally broad, with exponential tails and a large standard-deviation-to-mean ratio. Under several different perturbations the dynamics of the distribution is observed to be asymmetric, with a much longer time to build a wide expression distribution from below compared with a fast relaxation of the distribution toward steady state from above. These results show that the main features of the protein distributions are largely determined by population effects and are less sensitive to the intracellular biochemical noise.

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“…Here, we engineered a synthetic circuit that couples histidine synthesis to galactose utilization in the yeast Saccharomyces cerevisiae. The essential gene HIS3 was entirely deleted from its natural position and placed under GAL regulation ( Johnston and Carlson 1992;Lohr et al 1995;Jayadeva and Murthy 2001), together with a gfp reporter ( Figure 1a) (Li et al 2000;Braun and Brenner 2004). The recruitment of HIS3 to the GAL system presents serious challenges to the cell in histidine-lacking environments: in pure galactose, the GAL system must simultaneously support regulation of two metabolic tasks, galactose utilization and histidine biosynthesis.…”

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confidence: 99%

Synthetic Gene Recruitment Reveals Adaptive Reprogramming of Gene Regulation in Yeast

Stolovicki¹,

et al. 2006

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The recruitment of a gene to a foreign regulatory system is a major evolutionary event that can lead to novel phenotypes. However, the evolvability potential of cells depends on their ability to cope with challenges presented by gene recruitment. To study this ability, we combined synthetic gene recruitment with continuous culture and online measurements of the metabolic and regulatory dynamics over long timescales. The gene HIS3 from the histidine synthesis pathway was recruited to the GAL system, responsible for galactose utilization in the yeast S. cerevisiae. Following a switch from galactose to glucose-from induced to repressed conditions of the GAL system-in histidine-lacking chemostats (where the recruited HIS3 is essential), the regulatory system reprogrammed to adaptively tune HIS3 expression, allowing the cells to grow competitively in pure glucose. The adapted state was maintained for hundreds of generations in various environments. The timescales involved and the reproducibility of separate experiments render spontaneous mutations an unlikely underlying mechanism. Essentially all cells could adapt, excluding selection over a genetically variable population. The results reveal heritable adaptation induced by the exposure to glucose. They demonstrate that genetic regulatory networks have the potential to support highly demanding events of gene recruitment.

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Green fluorescent protein inSaccharomyces cerevisiae: Real-time studies of theGAL1 promoter

Cited by 77 publications

References 37 publications

Evolution of design principles in biochemical networks

Evolution of design principles in biochemical networks

Dynamics of protein distributions in cell populations

Synthetic Gene Recruitment Reveals Adaptive Reprogramming of Gene Regulation in Yeast

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