The Saccharomyces cerevisiae BAR1 gene encodes an exported protein with homology to pepsin.

MacKay, Vivian L.; Welch, Susan K.; Insley, Margaret Y.; Manney, Thomas R.; Holly, J; Saari, G.; Parker, Myra

doi:10.1073/pnas.85.1.55

Cited by 193 publications

(105 citation statements)

References 46 publications

(38 reference statements)

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“…The construction and use offkbl : : URA3 (Brizuela et al, 1991), cphl : : ADE2, cnal : : LEU2, cna2: : URA3 and cnbl : ; L YS2 (Foor et al, 1992) disruption fragments have been described. The bar1 : : ADE2 allele was constructed by synthesizing a BAR1 DNA fragment by polymerase chain reaction [residues 1 to 2729 of the sequence given by MacKay et al (1988)l and replacing the 1.4 kb XbaI-StuI fragment within the BAR1 gene with the 3.8 kb BamHI ADE2 fragment from p669 (provided by P. Linder; Foor ef al., 1992).…”

Section: Methodsmentioning

confidence: 99%

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Calcineurin-dependent growth of an FK506- and CsA-hypersensitive mutant of Saccharomyces cerevisiae

Parent

Nielsen²,

Morin³

et al. 1993

Journal of General Microbiology

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The immunosuppressants FK506 and cyclosporin A (CsA) bound to their receptors, FKBP12 or cyclophilin, inhibit the Ca2+/calmodulin-dependent protein phosphatase, calcineurin, preventing T cell activation or, in yeast, recovery from a-mating factor arrest. Vegetative growth of yeast does not require calcineurin, and in strains sensitive to FK506 or CsA, growth is inhibited by concentrations of drug much higher than those required to inhibit T cell activation or recovery from mating factor arrest. We now describe the isolation of a mutant of Saccharomyces cerevisiae which is 100-1000-fold more sensitive to the growth inhibitory properties of these drugs. The mutation @sI) also confers a slow growth phenotype which is partially suppressed by exogenously added Ca2+ and exacerbated by EGTA. Simultaneous disruption of the two genes (CNAI and CNA2) encoding the alternative forms of the catalytic A subunit of calcineurin, or of the gene (CNBI) encoding the regulatory B subunit, is lethal in anfksl mutant. Disruption of the gene encoding FKBP12 (FKBI) or the major, cytosolic cyclophilin (CPHI) infksl cells results in the loss of hypersensitivity to the relevant drug. Overexpression of CNAI or CNAZ, in conjunction with CNBI, results in a significant decrease in hypersensitivity to FK506 and CsA. The results show that the hypersensitivity of the fksI mutant is due to the inhibition of calcineurin phosphatase activity by the receptor-drug complexes. The growth dependence of the mutant on the Ca2+/calcineurin signal pathway provides an important tool for studying in yeast certain aspects of immune suppression by these drugs.

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Section: Methodsmentioning

confidence: 99%

“…However, the mutant and the wild-type strains did not differ in their response to a-factor (data not shown). To test this further, we disrupted BARI, the gene encoding a-factor protease (MacKay et al, 1988). Mutations in this gene confer hypersensitivity to a-factor.…”

Section: Expression Offksl-i and Mating Type Controlmentioning

confidence: 99%

Calcineurin-dependent growth of an FK506- and CsA-hypersensitive mutant of Saccharomyces cerevisiae

Parent

Nielsen²,

Morin³

et al. 1993

Journal of General Microbiology

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“…We then tested the possibility that other a-specific genes could also be expressed by performing semi-quantitative RT-PCR analysis, using as template RNA isolated from wild-type and itc1 strains of both mating types. Among the a-specific genes, we analysed STE2, the gene encoding the pheromone a-factor receptor (Blumer et al, 1988;Jenness et al, 1983); BAR1, encoding the protease that degrades a-factor (barrier protease) (Mackay et al, 1988); and ASG7, which has been recently shown to be involved in the regulation of the zygotic transition to vegetative growth (Roth et al, 2000). As shown in Fig.…”

Section: A-specific Genes Are Derepressed In the Mata Itc1 Mutantmentioning

confidence: 99%

Cell-type-dependent repression of yeast a-specific genes requires Itc1p, a subunit of the Isw2p–Itc1p chromatin remodelling complex

et al. 2003

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In Saccharomyces cerevisiae MATa haploid cells, the a-specific genes are expressed, whereas in the MATa haploid and MATa/a diploid cell types their transcription is repressed. It is shown in this report that Itc1p, a component of the ATP-dependent Isw2p-Itc1p chromatin remodelling complex, is required for the repression of a-specific genes. It has previously been reported that disruption of the ITC1 gene leads, in MATa cells, to an aberrant cell morphology resembling the polarized mating projection of cells responding to pheromone. The activation of the pheromone signalling pathway in itc1 mutants of both mating types was examined and found to be constitutively active in MATa itc1 but not in MATa itc1 cells. Furthermore, unlike the wild-type, MATa itc1 and MATa/a itc1/itc1 cells secrete a-factor and express significant levels of other a-specific genes. The results indicate that the inappropriate a-factor production in a MATa context, due to the derepression of the a-specific genes, produces an autocrine signalling loop that leads to the aberrant morphology displayed by MATa itc1 cells. It is suggested that the Isw2p-Itc1p complex contributes to maintain the repressive chromatin structure described for the asg operator present in the promoters of a-specific genes.

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“…Although pheromone secretion and detection are the essential elements for mating, additional, mating-type-specific genes make mating more efficient. One of these is the MATa-specific a-factor protease, Bar1 (Sprague and Herskowitz 1981;MacKay et al 1988), which helps MATa cells detect an a-factor gradient and polarize toward MATa partners (Jackson and Hartwell 1990; Barkai et al 1998). Yeast cells also express mating-type-specific agglutinins, which help cells attach to mating partners (Cappellaro et al 1991) in liquid but individually have little effect on mating efficiency on solid media (Lipke et al 1989;Roy et al 1991;de Nobel et al 1995).…”

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confidence: 99%

Genetically Engineered Transvestites Reveal Novel Mating Genes in Budding Yeast

Huberman

Murray

2013

Genetics

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Haploid budding yeast has two mating types, defined by the alleles of the MAT locus, MATa and MATa. Two haploid cells of opposite mating types mate by signaling to each other using reciprocal pheromones and receptors, polarizing and growing toward each other, and eventually fusing to form a single diploid cell. The pheromones and receptors are necessary and sufficient to define a mating type, but other mating-type-specific proteins make mating more efficient. We examined the role of these proteins by genetically engineering "transvestite" cells that swap the pheromone, pheromone receptor, and pheromone processing factors of one mating type for another. These cells mate with each other, but their mating is inefficient. By characterizing their mating defects and examining their transcriptomes, we found Afb1 (a-factor barrier), a novel MATa-specific protein that interferes with a-factor, the pheromone secreted by MATa cells. Strong pheromone secretion is essential for efficient mating, and the weak mating of transvestites can be improved by boosting their pheromone production. Synthetic biology can characterize the factors that control efficiency in biological processes. In yeast, selection for increased mating efficiency is likely to have continually boosted pheromone levels and the ability to discriminate between partners who make more and less pheromone. This discrimination comes at a cost: weak mating in situations where all potential partners make less pheromone. BIOLOGICAL processes are typically defined by the genes that are necessary and sufficient for function. However, in many cases, this minimal gene set does not encompass all the proteins involved in a process, and additional proteins promote biological efficiency. Finding these additional proteins may require the detection of subtle phenotypes, making it hard to know if all the genes involved in a process have been identified. One way to answer this question is to reengineer a pathway and ask whether the synthetic version fully mimics the natural function. Here, we show that this form of synthetic biology illuminates how cells of the budding yeast Saccharomyces cerevisiae mate efficiently.Budding yeast can be stably maintained as haploids or diploids. Haploids mate when two cells of opposite mating types signal to each other using reciprocal pheromones and receptors, polarize and grow toward each other, and eventually fuse to form a single diploid. Yeast has two mating types, a and a ( Figure 1A), determined by two alternative alleles at the MAT locus, MATa and MATa, which encode different transcription factors (Herskowitz 1988). These factors regulate the expression of mating-type-specific genes, many of which are involved with the production and detection of the pheromones that yeast cells use to signal to one another. The pheromones (a-and a-factor) are detected by G-proteincoupled receptors. MATa cells express a-factor (Betz and Duntze 1979), which is secreted through an ATP binding cassette transporter (Ste6) (McGrath and Varshavsky 1989)...

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The Saccharomyces cerevisiae BAR1 gene encodes an exported protein with homology to pepsin.

Cited by 193 publications

References 46 publications

Calcineurin-dependent growth of an FK506- and CsA-hypersensitive mutant of Saccharomyces cerevisiae

Calcineurin-dependent growth of an FK506- and CsA-hypersensitive mutant of Saccharomyces cerevisiae

Cell-type-dependent repression of yeast a-specific genes requires Itc1p, a subunit of the Isw2p–Itc1p chromatin remodelling complex

Genetically Engineered Transvestites Reveal Novel Mating Genes in Budding Yeast

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