Human G protein-coupled receptor studies in Saccharomyces cerevisiae

Liu, Rongfang; Wong, W. Y.; IJzerman, Adriaan P.

doi:10.1016/j.bcp.2016.02.010

Cited by 23 publications

(23 citation statements)

References 79 publications

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“…The eukaryotic, unicellular, budding yeast, Saccharomyces cerevisiae , is a useful microbial host organism for making GPCR biosensors . Compared with mammalian cell lines, S. cerevisiae provides a simple and predictable system for studying GPCR signaling because haploid S. cerevisiae expresses one type of GPCR (either the a ‐cell‐specific pheromone receptor, Ste2p, or the α‐cell‐specific pheromone receptor, Ste3p) and one type of heterotrimeric complex of G‐proteins (yeast Gα, Gβ, and Gγ; Gpa1p, Ste4p, and Ste18p, respectively).…”

Section: Introductionmentioning

confidence: 99%

Modifying Expression Modes of Human Neurotensin Receptor Type 1 Alters Sensing Capabilities for Agonists in Yeast Signaling Biosensor

Hashi

Nakamura

Ishii

et al. 2017

Biotechnology Journal

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Neurotensin receptor type 1 (NTSR1), a member of the G-protein-coupled receptor (GPCR) family, is naturally activated by binding of a neurotensin peptide, leading to a variety of physiological effects. The budding yeast Saccharomyces cerevisiae is a proven host organism for assaying the agonistic activation of human GPCRs. Previous studies showed that yeast cells can functionally express human NTSR1 receptor, permitting the detection of neurotensin-promoted signaling using a ZsGreen fluorescent reporter gene. However, the fluorescence intensity (sensitivity) of NTSR1-expressing yeast cells is low compared to that of yeast cells expressing other human GPCRs (e.g., human somatostatin receptors). The present study sought to increase the sensitivity of the NTSR1-expressing yeast for use as a fluorescent biosensor, including modification of the expression of human NTSR1 in yeast. Changes in the transcription, translation, and transport of the receptor are attempted by altering the promoter, consensus Kozak-like sequence, and secretion signal sequences of the NTSR1-encoding gene. The resulting yeast cells exhibited increased sensitivity to exogenously added peptide. The cells are further engineered by using cell-surface display technology to ensure that the agonistic peptides are secreted and tethered to the yeast cell wall, yielding cells with enhanced NTSR1 activation. This yeast biosensor holds promise for the identification of agonists to treat NTSR1-related diseases.

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Section: Introductionmentioning

confidence: 99%

Modifying Expression Modes of Human Neurotensin Receptor Type 1 Alters Sensing Capabilities for Agonists in Yeast Signaling Biosensor

Hashi

Nakamura

Ishii

et al. 2017

Biotechnology Journal

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“…Synthetic biology approaches, where human protein function and interactions are quantified by yeast-based gene circuits, have enabled high-throughput experiments at an impressive scale (Starita et al 2015;Sokolina et al 2017;Weile et al 2017;Woodsmith et al 2017). With genetic engineering, human GPCRs can be functionally linked to the yeast mating pathway, and mating-responsive reporter genes have allowed for detailed studies of GPCR activation (Liu et al 2016). As human GPCRs retain their natural preferences for ligands and G proteins in yeast (Brown et al 2000), this application of synthetic biology combines the highthroughput capabilities of yeast-based studies with the ability to rapidly characterize GPCR function in a cellular context.…”

mentioning

confidence: 99%

“…Despite the power of yeast-based studies of human GPCRs, only a small proportion of GPCRs have been functionally linked to the yeast mating pathway, and all have been ligand-activated (Liu et al 2016). Unlike these GPCRs, rhodopsin exists as a covalent complex between its lightsensitive chromophore 11-cis retinal and the seven-helix transmembrane opsin apoprotein (Smith 2010).…”

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confidence: 99%

Coupling of Human Rhodopsin to a Yeast Signaling Pathway Enables Characterization of Mutations Associated with Retinal Disease

et al. 2018

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G protein-coupled receptors (GPCRs) are crucial sensors of extracellular signals in eukaryotes, with multiple GPCR mutations linked to human diseases. With the growing number of sequenced human genomes, determining the pathogenicity of a mutation is challenging, but can be aided by a direct measurement of GPCR-mediated signaling. This is particularly difficult for the visual pigment rhodopsin-a GPCR activated by light-for which hundreds of mutations have been linked to inherited degenerative retinal diseases such as retinitis pigmentosa. In this study, we successfully engineered, for the first time, activation by human rhodopsin of the yeast mating pathway, resulting in signaling via a fluorescent reporter. We combine this novel assay for rhodopsin lightdependent activation with studies of subcellular localization, and the upregulation of the unfolded protein response in response to misfolded rhodopsin protein. We use these assays to characterize a panel of rhodopsin mutations with known molecular phenotypes, finding that rhodopsin maintains a similar molecular phenotype in yeast, with some interesting differences. Furthermore, we compare our assays in yeast with clinical phenotypes from patients with novel disease-linked mutations. We demonstrate that our engineered yeast strain can be useful in rhodopsin mutant classification, and in helping to determine the molecular mechanisms underlying their pathogenicity. This approach may also be applied to better understand the clinical relevance of other human GPCR mutations, furthering the use of yeast as a tool for investigating molecular mechanisms relevant to human disease.

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“…В целом можно сказать, что сигнальные пути с участием гетеротримерных G-белков у грибов структурно и функционально ближе к таковым у животных. Не случайно в дрожжах хорошо разработана система изучения GPCR и соответствующих G-белков человека с целью изучения лекарственных веществ [45].…”

Section: передача сигнала от гетеротримерного G-белка у животных и грunclassified

Role of the plant heterotrimeric G-proteins in the signal pathways regulation

Bovin

Долгих

2019

Ecological genetics

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Animal and fungal heterotrimeric G-proteins are among the well-known regulators of signaling pathways. Plant studies have shown that G-proteins may also be involved in the regulation of many processes. G-proteins are involved in hormonal regulation, control of cell proliferation, response to abiotic factors, control of biotic interactions and many others. It turned out that with a smaller variety of subunits, G-proteins of plants can have a greater variety of mechanisms for activating and transmitting signals. However, for most processes in plants the mechanisms of operation of heterotrimeric G-proteins remain poorly understood. This review is devoted to the analysis of modern ideas about the structure and functioning of heterotrimeric plant G proteins.

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Human G protein-coupled receptor studies in Saccharomyces cerevisiae

Cited by 23 publications

References 79 publications

Modifying Expression Modes of Human Neurotensin Receptor Type 1 Alters Sensing Capabilities for Agonists in Yeast Signaling Biosensor

Modifying Expression Modes of Human Neurotensin Receptor Type 1 Alters Sensing Capabilities for Agonists in Yeast Signaling Biosensor

Coupling of Human Rhodopsin to a Yeast Signaling Pathway Enables Characterization of Mutations Associated with Retinal Disease

Role of the plant heterotrimeric G-proteins in the signal pathways regulation

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