Synthesis of specific identified, phosphorylated, heat shock, and heat stroke proteins through the cell cycle of Saccharomyces cerevisiae.

Ludwig, J R; Foy, J J; Elliott, Steven; McLaughlin, Calvin S.

doi:10.1128/mcb.2.2.117

Cited by 59 publications

(36 citation statements)

References 51 publications

(30 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previous studies with S. cerevisiae have confirmed the cell cycle modulation of histones (20) but failed to turn up any other case of cell cycleperiodic protein synthesis (8)(9)(10)22); it might be expected that at least some of the cell cycleregulated proteins which we found should have been seen previously. There is no unitary resolution of this apparent discrepancy: each protein must be examined individually.…”

Section: Resultssupporting

confidence: 64%

“…Protein 6s was only slightly inhibited by temperature upshift and was not previously classified as a major heat shock protein, although it was substantially inhibited as a result of the manipulations inherent in the size fractionation procedure. DISCUSSION Although there have been reports of changes in many enzyme activities during the eucaryotic cell cycle (14,21,27; J. G. Yarger et al, in press), recent studies with S. cerevisiae (8)(9)(10)22) detected no changes in the synthesis of individual polypeptides and only two cases of cell cycle-periodic modification. Only histone synthesis was shown to be cell cycle regulated (20,30) in this earlier work.…”

Section: Resultsmentioning

confidence: 96%

“…cerevisiae proteins in the cell cycle (8)(9)(10)22), almost 200 proteins were examined quantitatively after being separated on two-dimensional (2- D) gels. The use of a double labeling method allowed the relative rates of synthesis of individual proteins to be measured by cutting the spots from gels and determining radioisotope ratios.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Identification of Proteins Whose Synthesis Is Modulated During the Cell Cycle ofSaccharomyces cerevisiae

Lörincz¹,

Miller²,

Xuong³

et al. 1982

Mol. Cell. Biol.

View full text Add to dashboard Cite

We examined the synthesis and turnover of individual proteins in the Saccharomyces cerevisiae cell cycle. Proteins were pulse-labeled with radioactive isotope (32S or 14C) in cells at discrete cycle stages and then resolved on two-dimensional gels and analyzed by a semiautomatic procedure for quantitating gel electropherogram-autoradiographs. The cells were obtained by one of three methods: (i) isolation of synchronous subpopulations of growing cells by zonal centrifugation; (ii) fractionation of pulse-labeled steady-state cultures according to cell age; and (iii) synchronization of cells with the mating pheromone, a-factor. In confirmation of previous studies, we found that the histones H4, H2A, and H2B were synthesized almost exclusively in the late Gl and early S phases. In addition, we identified eight proteins whose rates of synthesis were modulated in the cell cycle, and nine proteins (of which five, which may well be related, were unstable, with half-lives of 10 to 15 min) that might be regulated in the cell cycle by periodic synthesis, modification, or degradation. Based on the time of maximal labeling in the cell cycle and on experiments with a-factor and hydroxyurea, we assigned the cell cycle proteins to two classes: proteins in class I were labeled principally in early Gl phase and at a late stage of the cycle, whereas those in class II were primarily synthesized at times ranging from late Gl to mid S phase. At least one major control point for the cell cycle proteins occurred between "start" and early S phase. A set of stress-responsive proteins was also identified and analyzed. The rates of synthesis of these proteins were affected by certain perturbations that resulted during selection of synchronous cell populations and by heat shock.Understanding the molecular basis of the many cellular events which occur at discrete stages of the progress of cells from one division to the next is the central focus of research on the cell cycle. Saccharomyces cerevisiae has been developed as a model organism for studying eucaryotic cell cycle control (15-17, 34, 43). Genetic analysis has generated some of the basic ideas about functional pathways in the cell cycle and also provided material for further biochemical studies.In this paper, we report on a search for cell cycle-regulated proteins in S. cerevisiae. In an earlier analysis of the patterns of synthesis of S.cerevisiae proteins in the cell cycle (8-10, 22), almost 200 proteins were examined quantitatively after being separated on two-dimensional (2-t Present address:

show abstract

Section: Resultssupporting

confidence: 64%

Section: Resultsmentioning

confidence: 96%

See 1 more Smart Citation

Identification of Proteins Whose Synthesis Is Modulated During the Cell Cycle ofSaccharomyces cerevisiae

Lörincz¹,

Miller²,

Xuong³

et al. 1982

Mol. Cell. Biol.

View full text Add to dashboard Cite

show abstract

“…To this end we examined the effect of the cell cycle position on the thermotolerance of S. cerevisiae in exponentially growing cultures by using the elutriator rotor to separate cells by size and cell cycle position (9). We also examined the effects on the cell cycle of a mild heat shock, during which the cells acquire thermotolerance (10, 13).…”

mentioning

confidence: 99%

Effect of cell cycle position on thermotolerance in Saccharomyces cerevisiae

Plesset¹,

Ludwig²,

Cox³

et al. 1987

J Bacteriol

Self Cite

106

View full text Add to dashboard Cite

We showed that the heat killing ,curve for exponentially growing Saccharomyces cerevisiae was biphasic. This suggests two populations of cells with different therpial killing characteristics. Whep exponentially growing cells separated into cell cycle-specific fractions via centrifugal elutriation were, heat shocked, the fractions enriched in smnall unbudded cells showed greater resistance to heat killing than did other cell cycle fractions. The lethal effects of extreme temperature on Saccharomyces cerevisiae have been described. Exponentially growing cells are much more sensitive than are stationary-phase cells to a thermal shock (15). Thus, resting or quiescent cells are more thermotolerant than growing cells are. This heat resistance is acquired as the cells pass from the exponentially growing to the stationary state (12). Schenberg-Frascino and Moustacchi (15) isolated small unbudded cells from stationary cultures and observed their thermotolerance as they progressed through the first cell cycle into exponential growth. They observed that thermotolerance is decreased in budding cells. Yeast cells, when starved for an essential nutrient such as nitrogen, phosphorous, or sulfur, cease dividing and arrest their cell division as unbudded cells. The thermotolerance of cells starved for any of these nutrients resembles that of stationary-phase cells, and it has been suggested that this thermotolerance is a general characteristic of resting cells (11).Our aim was to clarify the relationship between the cell cycle and thermotolerance. To this end we examined the effect of the cell cycle position on the thermotolerance of S. cerevisiae in exponentially growing cultures by using the elutriator rotor to separate cells by size and cell cycle position (9). We also examined the effects on the cell cycle of a mild heat shock, during which the cells acquire thermotolerance (10, 13). In addition, we used several methods to arrest cells in the unbudded state, and we determined the thermotolerance of the resulting populations. The methods included arrest via deprivation of essential nutrients, alphafactor arrest, and the use of temperature-sensitive cell division cycle mutants. Our results demonstrate that a distinct population of unbudded cells was more thermotolerant than were cells in other morphological stages in the cell cycle. We suggest that this thermotolerance is associated with a defined state, G0, which has synthetic and physiological properties distinct from those of other unbudded cells. MATERIALS AND METHODSStrains. The following strains were used: SKQ2n (a/a adell+ +/ade2 +Ihisl); A364A (a lys2 tyrl his7 gall adel ade2 ural); cdc7-1 (ts124); cdc25-1 (ts321); cdc33-1 (El7a), derived from A364A (4; provided by L. Hartwell); cdc35-1 (BR214-4a; a adel ural his7 arg4 trpl); and cdc36-12 (ST21; a met2 cyh2) (provided by L. Hartwell).Media and cell growth.

show abstract

“…cerevisiae according to the phase of the cell cycle. Advantages and limitations of cell-size fractionation by means of this technique have been evaluated by several authors (Gordon & Elliott, 1977;Ludwig et al, 1982;Barford, 1986;Wheals, 1987;Van Doorn et al, 1988b;Mitchison, 1988). When the various fractions from the elutriator rotor are assigned to particular stages in the cell cycle, the variation in cell-size at any particular cell cycle stage, and the different sizes of mother and daughter cells (Hartwell, 1970;Carter & Jagadish, 1978) must be taken into consideration.…”

Section: Discussionmentioning

confidence: 99%

Changes in the Incorporation of Carbon Derived from Glucose into Cellular Pools During the Cell Cycle of Saccharomyces Cerevisiae

Oehlen

Doorn

Scholte

et al. 1990

Journal of General Microbiology

View full text Add to dashboard Cite

Synthesis of specific identified, phosphorylated, heat shock, and heat stroke proteins through the cell cycle of Saccharomyces cerevisiae.

Cited by 59 publications

References 51 publications

Identification of Proteins Whose Synthesis Is Modulated During the Cell Cycle ofSaccharomyces cerevisiae

Identification of Proteins Whose Synthesis Is Modulated During the Cell Cycle ofSaccharomyces cerevisiae

Effect of cell cycle position on thermotolerance in Saccharomyces cerevisiae

Changes in the Incorporation of Carbon Derived from Glucose into Cellular Pools During the Cell Cycle of Saccharomyces Cerevisiae

Contact Info

Product

Resources

About