Quantification of Yeast Flocculation

Soares, Eduardo V.; Mota, M.

doi:10.1002/j.2050-0416.1997.tb00940.x

Cited by 32 publications

(25 citation statements)

References 49 publications

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“…In the cell cycle, cbf12 expression peaks during stationary phase, possibly to allow flocculation to occur as a stress response under unfavorable conditions (29). We had noted that zfs1⌬ mutants often flocculated during stationary phase, a finding that was confirmed in the present study in a calcium-induced flocculation assay (35). We found that the disruption of cbf12 abolished the calcium-induced flocculation observed in zfs1⌬ mutants; this is therefore likely to be due to the intermediacy of Cbf12p, as shown by results with the double zfs1⌬ cbf12⌬ mutant.…”

Section: Discussionsupporting

confidence: 77%

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Posttranscriptional Regulation of Cell-Cell Interaction Protein-Encoding Transcripts by Zfs1p in Schizosaccharomyces pombe

Wells

Huang²,

Li³

et al. 2012

Molecular and Cellular Biology

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f Members of the tristetraprolin (TTP) family of CCCH tandem zinc finger proteins can bind directly to AU-rich elements in mRNAs and promote transcript deadenylation and decay. The yeast Schizosaccharomyces pombe expresses a single TTP family member, Zfs1p. In this study, we identified probable Zfs1p target mRNAs by comparing transcript levels in wild-type yeast and zfs1⌬ mutants, using deep sequencing and microarray approaches. We also used direct RNA sequencing to determine polyadenylation site locations and to confirm the presence of potential Zfs1p target sequences within the target mRNA. These studies identified a set of transcripts containing potential Zfs1p binding sites that accumulated significantly in the zfs1⌬ mutants; a subset of these transcripts decayed more slowly in the zfs1⌬ mutants and bound directly to Zfs1p in coimmunoprecipitation assays. One apparent direct target encodes the transcription factor Cbf12p, which is known to increase cell-cell adhesion and flocculation when overexpressed. Studies of zfs1⌬ cbf12⌬ double mutants demonstrated that the increased flocculation seen in zfs1⌬ mutants is due, at least in part, to a direct effect on the turnover of cbf12 mRNA. These data suggest that Zfs1p can both directly and indirectly regulate the levels of transcripts involved in cell-cell adhesion in this species.

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Section: Discussionsupporting

confidence: 77%

“…Flocculation in mid-log-phase yeast cells was measured as described previously (35). Briefly, 5 ml of logarithmically growing culture was centrifuged at 4,500 ϫ g for 5 min at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Posttranscriptional Regulation of Cell-Cell Interaction Protein-Encoding Transcripts by Zfs1p in Schizosaccharomyces pombe

Wells

Huang²,

Li³

et al. 2012

Molecular and Cellular Biology

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“…In anaerobic experiments, the wort was aerated only once prior to pitching and the fermentation was performed without aeration. The yeast flocculation ability was examined with the modified Stratford flocculation test and the pre-controlled micro sedimentation test 19 . Flow-cytometric analysis was performed with a PAS III from Partec (Münster, Germany)…”

Section: Methodsmentioning

confidence: 99%

Monitoring of Brewing Yeast Propagation Under Aerobic and Anaerobic Conditions Employing Flow Cytometry

Novák

Basařová

Teixeira

et al. 2007

Journal of the Institute of Brewing

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The vitality and viability of industrial strains of Saccharomyces cerevisiae was monitored during pilot plant experiments simulating yeast propagation under aerobic and anaerobic conditions. Industrial wort of 12°P original gravity was used as a growth substrate for yeast propagation. The work was carried out with three widely used Czech lager yeast industrial strains: strains 2, 7 and 95. Cell cycle, cell size, granularity, glycogen content, DNA and protein content were analyzed by flow cytometry. Significantly higher specific growth rates, higher content of yeast glycogen, earlier G2 /M phase cells maximum, and faster cell protein creation was observed under aerobic conditions compared to anaerobic. Strains 7 and 95 showed losses in flocculation ability after aerobic propagation compared to anaerobic propagation. Under either aerobic or strictly anaerobic conditions, only strain 2 did not show a significant loss in flocculation ability.

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“…We counted the number of cells in tightly clustered aggregates (flocs) out of 200 cells and subsequently performed a flocculation test to determine if cellular aggregates observed under the microscope exhibited rapid sedimentation characteristic of flocculent strains (Soares and Mota 1997). A flocculation rate assay described by Smit et al (1992) was modified for wild strains (Smit et al 1992).…”

Section: Genetic Analysismentioning

confidence: 99%

Effect of Domestication on the Spread of the [PIN+] Prion in Saccharomyces cerevisiae

2014

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Prions (infectious proteins) cause fatal neurodegenerative diseases in mammals. In the yeast Saccharomyces cerevisiae, many toxic and lethal variants of the [PSI+] and [URE3] prions have been identified in laboratory strains, although some commonly studied variants do not seem to impair cell growth. Phylogenetic analysis has revealed four major clades of S. cerevisiae that share histories of two prion proteins and largely correspond to different ecological niches of yeast. The [PIN+] prion was most prevalent in commercialized niches, infrequent among wine/vineyard strains, and not observed in ancestral isolates. As previously reported, the [PSI+] and [URE3] prions are not found in any of these strains. Patterns of heterozygosity revealed genetic mosaicism and indicated extensive outcrossing among divergent strains in commercialized environments. In contrast, ancestral isolates were all homozygous and wine/vineyard strains were closely related to each other and largely homozygous. Cellular growth patterns were highly variable within and among clades, although ancestral isolates were the most efficient sporulators and domesticated strains showed greater tendencies for flocculation.[PIN+]-infected strains had a significantly higher likelihood of polyploidy, showed a higher propensity for flocculation compared to uninfected strains, and had higher sporulation efficiencies compared to domesticated, uninfected strains. Extensive phenotypic variability among strains from different environments suggests that S. cerevisiae is a niche generalist and that most wild strains are able to switch from asexual to sexual and from unicellular to multicellular growth in response to environmental conditions. Our data suggest that outbreeding and multicellular growth patterns adapted for domesticated environments are ecological risk factors for the [PIN+] prion in wild yeast. P RIONS cause a variety of transmissible spongiform encephalopathies (TSEs) in mammals, including scrapie in sheep, bovine spongiform encephalopathy (BSE) in cattle, chronic wasting disease (CWD) in cervids, and CreutzfeldtJakob disease (CJD) in humans. With all TSEs, endogenous prion protein (PrP) misfolds into an altered, usually proteaseresistant, isoform, termed "PrP res " (" res" for protease resistant). Several prions have been described in fungi (Wickner 1994), with many prion variants of the [PSI+] and [URE3] prions showing toxic or even lethal effects in Saccharomyces cerevisiae (McGlinchey et al. 2011). The [PSI+] prion of S. cerevisiae is formed from the cytosolic protein Sup35p. There are three distinct regions of Sup35p: a glutamine/ asparagine-rich N-terminal domain (amino acids 1-123) that is necessary and sufficient for prion formation (TerAvanesyan et al. 1994) and that facilitates deadenylation and decay of messenger RNA (Hosoda et al. 2003); a middle M domain (amino acids 124-253) of unknown function; and an essential C-terminal domain that functions during translation termination (reviewed by Wickner et al. 2004 (Derkatch et ...

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Quantification of Yeast Flocculation

Cited by 32 publications

References 49 publications

Posttranscriptional Regulation of Cell-Cell Interaction Protein-Encoding Transcripts by Zfs1p in Schizosaccharomyces pombe

Posttranscriptional Regulation of Cell-Cell Interaction Protein-Encoding Transcripts by Zfs1p in Schizosaccharomyces pombe

Monitoring of Brewing Yeast Propagation Under Aerobic and Anaerobic Conditions Employing Flow Cytometry

Effect of Domestication on the Spread of the [PIN+] Prion in Saccharomyces cerevisiae

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