scholarly journals Oscillatory nucleocytoplasmic shuttling of the general stress response transcriptional activators Msn2 and Msn4 in Saccharomyces cerevisiae

2003 ◽  
Vol 161 (3) ◽  
pp. 497-505 ◽  
Author(s):  
Michel Jacquet ◽  
Georges Renault ◽  
Sylvie Lallet ◽  
Jan De Mey ◽  
Albert Goldbeter
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Single Cells ◽  
Time Lapse ◽  
Oscillatory Behavior ◽  
Transcriptional Activators ◽  
Nucleocytoplasmic Shuttling ◽  
Yeast Saccharomyces Cerevisiae ◽  
Nutritional Limitation ◽  
Response To Stress ◽  
General Response

Msn2 and Msn4 are two related transcriptional activators that mediate a general response to stress in yeast Saccharomyces cerevisiae by eliciting the expression of specific sets of genes. In response to stress or nutritional limitation, Msn2 and Msn4 migrate from the cytoplasm to the nucleus. Using GFP-tagged constructs and high-resolution time-lapse video microscopy on single cells, we show that light emitted by the microscope also triggers this migration. Unexpectedly, the population of Msn2 or Msn4 molecules shuttles repetitively into and out of the nucleus with a periodicity of a few minutes. A large heterogeneity in the oscillatory response to stress is observed between individual cells. This periodic behavior, which can be induced by various types of stress, at intermediate stress levels, is not dependent upon protein synthesis and persists when the DNA-binding domain of Msn2 is removed. The cAMP–PKA pathway controls the sensitivity of the oscillatory nucleocytoplasmic shuttling. In the absence of PKA, Msn4 continues to oscillate while Msn2 is maintained in the nucleus. We show that a computational model based on the possibility that Msn2 and Msn4 participate in autoregulatory loops controlling their subcellular localization can account for the oscillatory behavior of the two transcription factors.

scholarly journals The GTS1 gene, which contains a Gly-Thr repeat, affects the timing of budding and cell size of the yeast Saccharomyces cerevisiae.

1994 ◽  
Vol 14 (8) ◽  
pp. 5569-5578 ◽  
Author(s):  
K Mitsui ◽  
S Yaguchi ◽  
K Tsurugi
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acid ◽  
Cell Size ◽  
Single Cells ◽  
Exponential Growth Phase ◽  
Restrictive Temperature ◽  
Multicopy Plasmid ◽  
Wild Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
Mean Cell Volume

A gene with an open reading frame encoding a protein of 417 amino acid residues with a Gly-Thr repeat was isolated from the yeast Saccharomyces cerevisiae by using synthetic oligonucleotides encoding three Gly-Thr dimers as probes. The deduced amino acid sequence showed partial homology to the clock-affecting gene, per, of Drosophila melanogaster in the regions including the GT repeat. The function of the gene, named GTS1, was examined by characterizing the phenotypes of transformants with different copy numbers of the GTS1 gene produced either by inactivating the GTS1 gene by gene disruption (TM delta gts1) or by transformation with multicopy plasmid pPER119 (TMpGTS1). They grew at similar rates during the exponential growth phase, but the lag phases were shorter for TM delta gts1 and longer for TMpGTS1 cells than that for the wild type. Analyses of their cell cycle parameters using synchronized cells revealed that the unbudding period changed as a function of gene dosage; that is, the periods of TM delta gts1 and TMpGTS1 were about 20% shorter and longer, respectively, than that of the wild-type. Another significant change in the transformants was detected in the distribution of the cell size. The mean cell volume of the TM delta gts1 cells in the unbudded period (single cells) was 27% smaller than that of single wild-type cells, whereas that of single TMpGTS1 cells was 48% larger. Furthermore, in the temperature-sensitive cdc4 mutant, the GTS1 gene affected the timing of budding at the restrictive temperature. Thus, the GTS1 gene product appears to modulate the timing of budding to obtain an appropriate cell size independent of the DNA replication cycle.

Use of atomic force microscopy (AFM) to explore cell wall properties and response to stress in the yeast Saccharomyces cerevisiae

2013 ◽  
Vol 59 (4) ◽  
pp. 187-196 ◽  
Author(s):  
Jean Marie Francois ◽  
Cécile Formosa ◽  
Marion Schiavone ◽  
Flavien Pillet ◽  
Hélène Martin-Yken ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Atomic Force Microscopy ◽  
Cell Wall ◽  
Yeast Saccharomyces Cerevisiae ◽  
Force Microscopy ◽  
Atomic Force ◽  
Wall Properties ◽  
Response To Stress ◽  
Cell Wall Properties

scholarly journals Symmetric cell division in pseudohyphae of the yeast Saccharomyces cerevisiae.

1994 ◽  
Vol 5 (9) ◽  
pp. 1003-1022 ◽  
Author(s):  
S J Kron ◽  
C A Styles ◽  
G R Fink
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Cell Size ◽  
Time Lapse ◽  
Cell Types ◽  
Yeast Saccharomyces Cerevisiae ◽  
Distinct Cell ◽  
Mother And Daughter ◽  
Dividing Cells ◽  
Unique Cell

Laboratory strains of Saccharomyces cerevisiae are dimorphic; in response to nitrogen starvation they switch from a yeast form (YF) to a filamentous pseudohyphal (PH) form. Time-lapse video microscopy of dividing cells reveals that YF and PH cells differ in their cell cycles and budding polarity. The YF cell cycle is controlled at the G1/S transition by the cell-size checkpoint Start. YF cells divide asymmetrically, producing small daughters from full-sized mothers. As a result, mothers and daughters bud asynchronously. Mothers bud immediately but daughters grow in G1 until they achieve a critical cell size. By contrast, PH cells divide symmetrically, restricting mitosis until the bud grows to the size of the mother. Thus, mother and daughter bud synchronously in the next cycle, without a G1 delay before Start. YF and PH cells also exhibit distinct bud-site selection patterns. YF cells are bipolar, producing their second and subsequent buds at either pole. PH cells are unipolar, producing their second and subsequent buds only from the end opposite the junction with their mother. We propose that in PH cells a G2 cell-size checkpoint delays mitosis until bud size reaches that of the mother cell. We conclude that yeast and PH forms are distinct cell types each with a unique cell cycle, budding pattern, and cell shape.

scholarly journals The glutamine-rich activation domains of human Sp1 do not stimulate transcription in Saccharomyces cerevisiae.

1995 ◽  
Vol 15 (2) ◽  
pp. 983-988 ◽  
Author(s):  
A S Ponticelli ◽  
T S Pardee ◽  
K Struhl
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Rna Polymerase Ii ◽  
Binding Protein ◽  
Tata Box ◽  
Transcriptional Activators ◽  
Yeast Saccharomyces Cerevisiae ◽  
Activation Domains ◽  
Tata Box Binding Protein ◽  
Simplex Virus ◽  
Rna Polymerase Ii Transcription

Eukaryotic transcriptional activators have been classified on the basis of the characteristics of their activation domains. Acidic activation domains, such as those in the yeast GAL4 or GNC4 proteins and the herpes simplex virus activator VP16, stimulate RNA polymerase II transcription when introduced into a variety of eukaryotic cells. This species interchangeability demonstrates that the mechanism by which acidic activation domains function is highly conserved in the eukaryotic kingdom. To determine whether such a conservation of function exists for a different class of activation domain, we have tested whether the glutamine-rich activation domains of the human transcriptional activator Sp1 function in the yeast Saccharomyces cerevisiae. We report here that the glutamine-rich domains of Sp1 do not stimulate transcription in S. cerevisiae, even when accompanied by human TATA-box binding protein (TBP) or human-yeast TATA-box binding protein hybrids. Thus, in contrast to the case for acidic activation domains, the mechanism by which glutamine-rich domains stimulate transcription is not conserved between S. cerevisiae and humans.

scholarly journals Cell cycle phase expansion in nitrogen-limited cultures of Saccharomyces cerevisiae.

1980 ◽  
Vol 85 (1) ◽  
pp. 96-107 ◽  
Author(s):  
C J Rivin ◽  
W L Fangman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Cycle Length ◽  
Time Lapse ◽  
S Phase ◽  
Cycle Phase ◽  
Yeast Saccharomyces Cerevisiae ◽  
Bud Emergence ◽  
Length Changes ◽  
Cell Cycle Length

The time and coordination of cell cycle events were examined in the budding yeast Saccharomyces cerevisiae. Whole-cell autoradiographic techniques and time-lapse photography were used to measure the duration of the S, G1, and G2 phases, and the cell cycle positions of "start" and bud emergence, in cells whose growth rates were determined by the source of nitrogen. It was observed that the G1, S, and G2 phases underwent a proportional expansion with increasing cell cycle length, with the S phase occupying the middle half of the cell cycle. In each growth condition, start appeared to correspond to the G1 phase/S phase boundary. Bud emergence did not occur until mid S phase. These results show that the rate of transit through all phases of the cell cycle can vary considerably when cell cycle length changes. When cells growing at different rates were arrested in G1, the following synchronous S phase were of the duration expected from the length of S in each asynchronous population. Cells transferred from a poor nitrogen source to a good one after arrest in G1 went through the subsequent S phase at a rate characteristic of the better medium, indicating that cells are not committed in G1 to an S phase of a particular duration.

scholarly journals Improved use of a public good selects for the evolution of undifferentiated multicellularity

eLife ◽  
2013 ◽  
Vol 2 ◽  
Author(s):  
John H Koschwanez ◽  
Kevin R Foster ◽  
Andrew W Murray
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Experimental Evolution ◽  
Cell Separation ◽  
Rational Design ◽  
Single Cells ◽  
Evolutionary Strategies ◽  
Hexose Transporter ◽  
Yeast Saccharomyces Cerevisiae ◽  
Know How ◽  
Transporter Expression

We do not know how or why multicellularity evolved. We used the budding yeast, Saccharomyces cerevisiae, to ask whether nutrients that must be digested extracellularly select for the evolution of undifferentiated multicellularity. Because yeast use invertase to hydrolyze sucrose extracellularly and import the resulting monosaccharides, single cells cannot grow at low cell and sucrose concentrations. Three engineered strategies overcame this problem: forming multicellular clumps, importing sucrose before hydrolysis, and increasing invertase expression. We evolved populations in low sucrose to ask which strategy they would adopt. Of 12 successful clones, 11 formed multicellular clumps through incomplete cell separation, 10 increased invertase expression, none imported sucrose, and 11 increased hexose transporter expression, a strategy we had not engineered. Identifying causal mutations revealed genes and pathways, which frequently contributed to the evolved phenotype. Our study shows that combining rational design with experimental evolution can help evaluate hypotheses about evolutionary strategies.

scholarly journals Patterns of bud-site selection in the yeast Saccharomyces cerevisiae.

1995 ◽  
Vol 129 (3) ◽  
pp. 751-765 ◽  
Author(s):  
J Chant ◽  
J R Pringle
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Time Lapse ◽  
Growth Conditions ◽  
Alpha Cells ◽  
Yeast Saccharomyces Cerevisiae ◽  
Cell Cycles ◽  
Daughter Cells ◽  
Division Site ◽  
Mother Cells ◽  
Bud Site

Cells of the yeast Saccharomyces cerevisiae select bud sites in either of two distinct spatial patterns, known as axial (expressed by a and alpha cells) and bipolar (expressed by a/alpha cells). Fluorescence, time-lapse, and scanning electron microscopy have been used to obtain more precise descriptions of these patterns. From these descriptions, we conclude that in the axial pattern, the new bud forms directly adjacent to the division site in daughter cells and directly adjacent to the immediately preceding division site (bud site) in mother cells, with little influence from earlier sites. Thus, the division site appears to be marked by a spatial signal(s) that specifies the location of the new bud site and is transient in that it only lasts from one budding event to the next. Consistent with this conclusion, starvation and refeeding of axially budding cells results in the formation of new buds at nonaxial sites. In contrast, in bipolar budding cells, both poles are specified persistently as potential bud sites, as shown by the observations that a pole remains competent for budding even after several generations of nonuse and that the poles continue to be used for budding after starvation and refeeding. It appears that the specification of the two poles as potential bud sites occurs before a daughter cell forms its first bud, as a daughter can form this bud near either pole. However, there is a bias towards use of the pole distal to the division site. The strength of this bias varies from strain to strain, is affected by growth conditions, and diminishes in successive cell cycles. The first bud that forms near the distal pole appears to form at the very tip of the cell, whereas the first bud that forms near the pole proximal to the original division site (as marked by the birth scar) is generally somewhat offset from the tip and adjacent to (or overlapping) the birth scar. Subsequent buds can form near either pole and appear almost always to be adjacent either to the birth scar or to a previous bud site. These observations suggest that the distal tip of the cell and each division site carry persistent signals that can direct the selection of a bud site in any subsequent cell cycle.

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