(A) Minute-long trajectories of the POA1 locus from both (+) glucose (blue) and (–) glucose (red) conditions projected on bright field images. Log-growing cells in (+) glucose were acutely starved …
(A) Individual log-log MSD plots of POA1 loci in non-starved (left) and starved (right) cells. (B) Individual log-log MSD plots of GFA1 mRNP particles in non-starved (left) and starved (right) …
(A) Log-log MSD plot of the URA3 locus during exponential growth and after acute starvation. (B) Log-log MSD plot of the GFA1 mRNP during exponential growth and quiescence (see 'Materials and …
(A) Quantification of filamentous actin during logarithmic growth, (+) glucose, and after acute starvation, (–) glucose. Cells were fixed and stained with phalloidin. Z-stack projections were then …
(A) Log-log MSD plot of the FBA1 mRNP after treatment as described in Figure 2C. (B) Log-log MSD plot of the URA3 locus after treatment as described in Figure 2C. (C) Log-log MSD plot of the pLacO …
(A) Intracellular ATP concentrations of acutely glucose-starved yeast were back-diluted into media containing 2% dextrose (n = 2 experiments), 2% dextrose + 0.02% azide (n = 2 experiments), or …
Log-growing wild-type (WT) and cbp2∆ yeast cells were acutely starved for glucose and back-diluted into media containing 2% dextrose (WT) or maintained in (–) glucose media. Intracellular ATP …
(A) Boxplots of the pHi of cells acutely starved for glucose or treated with varying concentrations of potassium sorbate (K+Sorbate). Log-growing yeast cells expressing phluorin (Miesenböck et al., …
(A) Intracellular pH calibration curve (Orij et al., 2009) (see 'Materials and methods'). Error bars represent SD. (B) Log-log MSD plot of the URA3 locus. Cells were treated as described in Figure 4A.
(A) Intracellular ATP concentrations after differing treatments with 0.02% sodium azide (‘wash’ versus ‘spike’) (see 'Materials and methods'). Intracellular ATP concentrations were determined as in F…
(A) Nuclear volume after acute glucose starvation. Histograms of nuclear volumes measured by reconstruction from three-dimensional image stacks using Imaris. The p-value resulting from a two-tailed …
(A) Cells were treated with K+Sorbate as in Figure 4 and cell volume measured as in Figure 5B. (B) Cells were starved of glucose in either low or high pH (5.0 or 7.4) and cell volume was measured.
(A) Intracellular ATP concentrations after treatment with either 0.4 M or 0.8 M NaCl. Cells were treated as described in Figure 5E and intracellular ATP concentrations were determined as in Figure 3A…
(A) Dry mass measurements of non-starved and starved yeast. Yeast cells in normal glucose, glucose deprivation, and raffinose growth conditions were lyophilized, and their cellular dry masses were …
Both glucose-starved and non-starved budding yeast cells show an anti-correlation of cell stiffness with cell diameter. Linear fitting R2 value for non-starved cell data = 0.54; R2 value for starved …
Intracellular ATP concentrations of log-growing yeast were determined as in Figure 3A after treatment with latrunculin A or the solvent control DMSO. The zero minute time point was taken immediately …
(A) MSD curves for GFA1 mRNP particles in non-starved cells (blue), cells acutely starved of glucose (red), and cells acutely starved of glucose additionally treated with 20 mM 2-deoxyglucose (2DG) …
Effective diffusion coefficients (K; µm2/s) and anomalous diffusion exponents (α) for macromolecules in each condition.
Condition | POA1 Locus | pLacO Plasmid | URA3 Locus | GFA1 mRNP | FBA1 mRNP | |||||
---|---|---|---|---|---|---|---|---|---|---|
K | α | K | α | K | α | K | α | K | α | |
(+) Glucose | 0.0057 | 0.69 | 0.0067 | 0.78 | 0.0076 | 0.65 | 0.0420 | 0.83 | 0.0501 | 0.85 |
(-) Glucose | 0.0023 | 0.64 | 0.0021 | 0.80 | 0.0022 | 0.73 | 0.0131 | 0.77 | 0.0139 | 0.77 |
(-) Glucose pH 7.4 | 0.0015 | 0.65 | -- | -- | -- | -- | 0.0120 | 0.75 | -- | -- |
DMSO | 0.0059 | 0.56 | 0.0046 | 0.70 | 0.0060 | 0.61 | 0.0491 | 0.83 | 0.0541 | 0.83 |
Nocodazole | 0.0040 | 0.48 | 0.0025 | 0.57 | 0.0046 | 0.51 | 0.0364 | 0.85 | 0.0397 | 0.85 |
Latrunculin A | 0.0038 | 0.50 | 0.0024 | 0.63 | 0.0038 | 0.55 | 0.0476 | 0.82 | 0.0550 | 0.81 |
Nocodazole + LatA | 0.0028 | 0.48 | 0.0014 | 0.49 | 0.0026 | 0.52 | 0.0303 | 0.81 | 0.0367 | 0.82 |
2 mM K+Sorbate | 0.0056 | 0.73 | 0.0051 | 0.80 | 0.0059 | 0.68 | 0.0402 | 0.82 | 0.0296 | 0.80 |
4 mM K+Sorbate | 0.0050 | 0.75 | 0.0044 | 0.72 | 0.0048 | 0.71 | 0.0406 | 0.78 | 0.0242 | 0.79 |
6 mM K+Sorbate | 0.0039 | 0.70 | 0.0018 | 0.66 | 0.0027 | 0.66 | 0.0378 | 0.76 | 0.0270 | 0.78 |
8 mM K+Sorbate | 0.0023 | 0.64 | 0.0012 | 0.61 | 0.0014 | 0.61 | 0.0340 | 0.76 | 0.0164 | 0.77 |
0.4 M NaCl | 0.0030 | 0.69 | 0.0026 | 0.75 | -- | -- | 0.0129 | 0.83 | 0.0146 | 0.85 |
0.6 M NaCl | 0.0012 | 0.60 | 0.0011 | 0.58 | -- | -- | 0.0047 | 0.83 | 0.0057 | 0.84 |
0.8 M NaCl | 0.0009 | 0.63 | 0.0011 | 0.63 | -- | -- | 0.0013 | 0.67 | 0.0016 | 0.77 |
Quiescence | -- | -- | -- | -- | -- | -- | 0.0004 | 0.29 | 0.0015 | 0.68 |
0.02% Azide (Wash) | 0.0037 | 0.74 | -- | -- | -- | -- | 0.0293 | 0.82 | -- | -- |
0.02% Azide (Spike) | 0.0012 | 0.67 | -- | -- | -- | -- | 0.0155 | 0.81 | -- | -- |
Yeast strains used in this study.
Strain | Genotype | Source |
---|---|---|
KWY165 | W303; MATa ura3-1 leu2-3 his3-11,15 trp1-1 ade2-1 | This Study |
KWY1622 | W303; MATα ybr022w::256LacO::LEU2 his3::LacI-GFP::HIS3 trp1::dsRED-HDEL::TRP1 | Green et al., 2012 |
KWY3541 | KWY 1622, cbp2∆::KANMX | This Study |
KWY3538 | W303; MATα his3::LacI-GFP::HIS3 trp1::dsRED-HDEL::TRP1 256LacO::URA3 | This Study |
KWY2848 | W303; MATα his3::LacI-GFP::HIS3 trp1::dsRED-HDEL::TRP1 yel021w::128LacO::URA3 | This Study |
KWY4586 | W303; MATa ybr022w::112TetO::URA3 yfr023w::256LacO::LEU2 his3::LacI-GFP_TetR-3XmCherry::HIS3 | This Study |
KWY970 | KWY 165, ura3::pHIS-GFP-TUB1::URA3 | This Study |
KWY3661 | W303; MATa pHluorin::URA3 | This Study |
KWY4796 | W303; MATα trp1::dsRED-HDEL::TRP1 leu2::TetR-GFP::LEU2 | This Study |
KWY4736 | W303 MATa/S288c MATα NDC1/ndc1::NDC1-tdTomato::KANMX GFA1/gfa1::GFA1-24PP7 3xYFP-PP7-CFP::HIS3 | This Study |
KWY4737 | W303 MATa/S288c MATα NDC1/ndc1::NDC1-tdTomato::KANMX FBA1/fba1::FBA1-24PP7 3xYFP-PP7-CFP::HIS3 | This Study |
KWY5112 | W303; MATa ura3-1 leu2-3 his3-11,15 trp1-1 ade2-1 VPH1::VPH1-mCherry::KANMX leu2::GFP::CaURA3 | This Study |
KWY6241 | CAF13 (S. pombe wildtype) | This Study |
yYB5978 | S288c; MATa his3∆1 leu2∆0 ura3∆0 met15∆0 LYS2 ADE2 TRP1 bar1::kanMX | Caudron and Barral, 2013 |
Plasmids used in this study.
Plasmid | Description | Source |
---|---|---|
pKW2734 | pHMX-256LacO (CEN, URA3) | This Study |
pKW2957 | pYES2-PACT1-pHluorin (CEN, URA3) | Orij et al., 2009 |
pKW544 | MET25pro-PP7-CP-3xYFP (CEN, HIS3) | This Study |