Disruption of six novel yeast genes located on chromosome II reveals one gene essential for vegetative growth and two required for sporulation and conferring hypersensitivity to various chemicals

Kucharczyk, Róża; Gromadka, Robert; Migdalski, Andrzej; Słonimski, Piotr P.; Rytka, Joanna

doi:10.1002/(sici)1097-0061(199907)15:10b<987::aid-yea403>3.0.co;2-5

Cited by 33 publications

(19 citation statements)

References 28 publications

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“…HSL7 and IRA1 are located only 13 kb apart from each other on chromosome II, indicating that this cosegregation could be due to linkage rather than the contribution of both genes to the trait. Although null mutations in HSL7 are associated with elongated cell morphology in other common strain backgrounds S1278B and W303 (Fujita et al 1999;Kucharczyk et al 1999), this morphological change is not observed in the S288C null mutant (Kucharczyk et al 1999), which we have confirmed using the yeast knockout collection (Giaever et al 2002). The S288C background of this evolved strain therefore makes IRA1 the more likely modifier, though the phenotype could also require both mutations.…”

Section: Phenotypic Variation Suggests Secondary Modifiers Influence supporting

confidence: 62%

Experimental evolution reveals favored adaptive routes to cell aggregation in yeast

Hope

Amorosi

Miller

et al. 2016

Preprint

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Yeast flocculation is a community-building cell aggregation trait that is an important mechanism of stress resistance and a useful phenotype for brewers; however, it is also a nuisance in many industrial processes, in clinical settings, and in the laboratory. Chemostat-based evolution experiments are impaired by inadvertent selection for aggregation, which we observe in 35% of populations. These populations provide a testing ground for understanding the breadth of genetic mechanisms Saccharomyces cerevisiae uses to flocculate, and which of those mechanisms provide the biggest adaptive advantages. In this study, we employed experimental evolution as a tool to ask whether one or many routes to flocculation are favored, and to engineer a strain with reduced flocculation potential. Using a combination of whole genome sequencing and bulk segregant analysis, we identified causal mutations in 23 independent clones that had evolved cell aggregation during hundreds of generations of chemostat growth. In 12 of those clones, we identified a transposable element insertion in the promoter region of known flocculation gene FLO1, and, in an additional five clones, we recovered loss-of-function mutations in transcriptional repressor TUP1, which regulates FLO1 and other related genes. Other causal mutations were found in genes that have not been previously connected to flocculation. Evolving a flo1 deletion strain revealed that this single deletion reduces flocculation occurrences to 3%, and demonstrated the efficacy of using experimental evolution as a tool to identify and eliminate the primary adaptive routes for undesirable traits.

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Section: Phenotypic Variation Suggests Secondary Modifiers Influence supporting

confidence: 62%

Experimental evolution reveals favored adaptive routes to cell aggregation in yeast

Hope

Amorosi

Miller

et al. 2016

Preprint

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“…Although mon1⌬ has not been previously reported as having a role in the Cvt pathway, complementation analyses indicate that CCZ1 is allelic with CVT16, a previously uncharacterized CVT gene (10). The ccz1⌬ mutant was originally identified due to its sensitivity to caffeine, calcium, and zinc (29). It has also been shown that the strain displays a severe vacuole protein-sorting defect.…”

Section: Mon1 and Ccz1mentioning

confidence: 99%

The Ccz1-Mon1 Protein Complex Is Required for the Late Step of Multiple Vacuole Delivery Pathways

Wang

Strømhaug

Shima

et al. 2002

Journal of Biological Chemistry

128

120

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Mon1 and Ccz1 were identified from a gene deletion library as mutants defective in the vacuolar import of aminopeptidase I (Ape1) via the cytoplasm to vacuole targeting (Cvt) pathway. The mon1⌬ and ccz1⌬ strains also displayed defects in autophagy and pexophagy, degradative pathways that share protein machinery and mechanistic features with the biosynthetic Cvt pathway. Further analyses indicated that Mon1, like Ccz1, was required in nearly all membrane-trafficking pathways where the vacuole represented the terminal acceptor compartment. Accordingly, both deletion strains had kinetic defects in the biosynthetic delivery of resident vacuolar hydrolases through the CPY, ALP, and MVB pathways. Biochemical and microscopy studies suggested that Mon1 and Ccz1 functioned after transport vesicle formation but before (or at) the fusion step with the vacuole. Thus, ccz1⌬ and mon1⌬ are the Compartmentalization allows eukaryotic cells to regulate intracellular functions by separating competing reactions and localizing enzymes and substrates at specific locations within the cell. Efficient compartmentalization necessitates dynamic protein trafficking processes by which cells are able to establish and maintain the identity and function of each organelle. The vacuole (lysosome) of the yeast Saccharomyces cerevisiae plays a central role in the turnover of cytoplasmic organelles, degradation of intracellular/extracellular components, and maintenance of cellular physiology (1). To carry out these functions, the vacuole maintains a variety of degradative enzymes. Both resident hydrolases and their substrates arrive at this destination through a variety of sorting pathways. The main routes by which vacuolar hydrolases are delivered to this organelle are the carboxypeptidase Y (CPY), 1 alkaline phosphatase (ALP), and multivesicular body (MVB) pathways, which involve transit through a portion of the secretory pathway, and the cytoplasm to vacuole targeting (Cvt) pathway by which the cargo molecules are packaged as cytosolic membrane-bound intermediates (2, 3). Resident proteins are also transmitted by inheritance from mother cell vacuoles to daughter cells during cell division (4). Substrates enter the vacuole through endocytosis, autophagy and the vacuole import and degradation pathway (reviewed in Ref. 5). One common feature in all of these processes is membrane fusion. The membrane fusion mechanism acts to ensure specificity for the directed movement of proteins while also maintaining the distinct composition of each organelle within the highly compartmentalized eukaryotic cell.The cytoplasm to vacuole targeting pathway that is used to deliver the soluble hydrolase aminopeptidase I (Ape1) to the vacuole has been under investigation (for reviews see Refs. 2, 5, and 6). Under vegetative conditions, precursor Ape1 (prApe1) is assembled into a large Cvt complex composed in part of multiple prApe1 dodecamers in the cytosol that becomes enwrapped within a double-membrane Cvt vesicle (7). Upon completion, the cytosolic Cvt vesicle target...

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“…It is well established that there is some variation among laboratory yeast strains of different genetic backgrounds reflected, for example, in differences between mutant or deletion phenotypes in different strains; at its extreme, genes reported as essential in a strain of one genetic background were shown to be dispensable in another (24,25). We checked whether two yeast strains, W303-1B and BY4742, widely used to obtain defined deletions of yeast open reading frames, display the same phenotype linked to the deletion of ZWF1 as described previously for other strains with presumably different genetic back- …”

Section: Ald6 and Zms1 Are Multicopy Suppressors Of Zwf1⌬ Strainmentioning

confidence: 99%

The ALD6 Gene Product Is Indispensable for Providing NADPH in Yeast Cells Lacking Glucose-6-phosphate Dehydrogenase Activity

Grabowska

Chełstowska

2003

Journal of Biological Chemistry

118

101

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Reducing equivalents in the form of NADPH are essential for many enzymatic steps involved in the biosynthesis of cellular macromolecules. An adequate level of NADPH is also required to protect cells against oxidative stress. The major enzymatic source of NADPH in the cell is the reaction catalyzed by glucose-6-phosphate dehydrogenase, the first enzyme in the pentose phosphate pathway. Disruption of the ZWF1 gene, encoding glucose-6-phosphate dehydrogenase in the yeast Saccharomyces cerevisiae, results in methionine auxotrophy and increased sensitivity to oxidizing agents. It is assumed that both phenotypes are due to an NADPH deficiency in the zwf1⌬ strain. We used a Met ؊ phenotype displayed by the zwf1⌬ strain to look for multicopy suppressors of this deletion. We found that overexpression of the ALD6 gene coding for cytosolic acetaldehyde dehydrogenase, which utilizes NADP ؉ as its cofactor, restores the Met ؉ phenotype of the zwf1⌬ strain. Another multicopy suppressor identified in our screen, the ZMS1 gene encoding a putative transcription factor, regulates the level of ALD6 expression. A strain bearing a double ZWF1 ALD6 gene disruption is not viable. Thus, our results indicate the reaction catalyzed by Ald6p as an important source of reducing equivalents in the yeast cells.

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Disruption of six novel yeast genes located on chromosome II reveals one gene essential for vegetative growth and two required for sporulation and conferring hypersensitivity to various chemicals

Cited by 33 publications

References 28 publications

Experimental evolution reveals favored adaptive routes to cell aggregation in yeast

Experimental evolution reveals favored adaptive routes to cell aggregation in yeast

The Ccz1-Mon1 Protein Complex Is Required for the Late Step of Multiple Vacuole Delivery Pathways

The ALD6 Gene Product Is Indispensable for Providing NADPH in Yeast Cells Lacking Glucose-6-phosphate Dehydrogenase Activity

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