Mitochondria reorganization upon proliferation arrest predicts individual yeast cell fate

  1. Damien Laporte
  2. Laëtitia Gouleme
  3. Laure Jimenez
  4. Ines Khemiri
  5. Isabelle Sagot  Is a corresponding author
  1. Centre National de la Recherche Scientifique, Université de Bordeaux - Institut de Biochimie et Génétique Cellulaires, France
11 figures, 1 table and 2 additional files

Figures

Figure 1 with 1 supplement
Mitochondrial network reorganization in non-proliferating cells.

(A) Mitochondrial network organization in WT cells expressing Ilv3-RFP as a function of time in YPD (OD600nm: circles). Cells with a tubular (violet), a fragmented (blue), a vesicular (green), a …

https://doi.org/10.7554/eLife.35685.002
Figure 1—source data 1

Cell category scoring for each replicated experiment in Figure 1 panel A, B and F to H.

https://doi.org/10.7554/eLife.35685.005
Figure 1—figure supplement 1
Mitochondrial network organization in WT cells experiencing an extended period of non-proliferation or upon starvation for various nutrients and localization of various mitochondrial proteins in non-proliferating cells.

(A) Proliferation in YPD or YPGE of WT cells of the indicated genotype monitored by measuring OD600nm as a function of time. (B) Mitochondrial network organization of WT cells expressing Ilv3-RFP …

https://doi.org/10.7554/eLife.35685.003
Figure 1—figure supplement 1—source data 1

Cell category scoring for each replicated experiment in Figure 1—figure supplement 1 panel A, B and F.

https://doi.org/10.7554/eLife.35685.004
Figure 2 with 1 supplement
Mitochondrial network reorganization can predict cell fate.

(A) WT cells expressing Ilv3-RFP were grown 7 d in YPD and stained with methylene blue to reveal dead cells. Cells surrounded by a white dashed line are methylene blue positive. The histogram shows …

https://doi.org/10.7554/eLife.35685.006
Figure 2—source data 1

Cell category scoring for each replicated experiment in Figure 2 panel A, B, C, D and E to G.

https://doi.org/10.7554/eLife.35685.009
Figure 2—figure supplement 1
Mitochondrial network organization in rho zero cells, ERMES mutant and WT cells of several genetic backgrounds upon proliferation cessation and quiescence exit.

(A) Upon quiescence exit, the mitochondrial network re-forms tubule before bud emergence. WT cells were grown for 7 days in YPD and allow to exit quiescence in new YPD liquid medium. The morphology …

https://doi.org/10.7554/eLife.35685.007
Figure 2—figure supplement 1—source data 1

Cell category scoring for each replicated experiment in Figure 2—figure supplement 1 panel A, C, E, F and G.

https://doi.org/10.7554/eLife.35685.008
Figure 3 with 1 supplement
Cell volume and individual cell fate.

(A) Cell volume distribution of WT cells grown for 7, 14, and 21 d in YPD. In A to C, red dashed lines define three sub-populations each representing 1/3rd of the total population. Dead cells (black …

https://doi.org/10.7554/eLife.35685.010
Figure 3—source data 1

Cell category scoring for each replicated experiment in Figure 3 panel A, B, and C.

https://doi.org/10.7554/eLife.35685.013
Figure 3—figure supplement 1
Cell viability and quiescence exit capacity after proliferation cessation in function of cell volume.

(A) WT prototroph cells grown for the indicated time in YPD were individually separated by micro-manipulation onto YPD plates according to their volume (n = 120, N = 2). Representative images of the …

https://doi.org/10.7554/eLife.35685.011
Figure 3—figure supplement 1—source data 1

Cell category scoring for each replicated experiment in Figure 3—figure supplement 1 panel A and B.

https://doi.org/10.7554/eLife.35685.012
The capacity to enter quiescence does not correlate with a cell’s replicative age or density.

(A–D) WT cells expressing Ilv3-RFP were grown 7 d in YPD, stained with calcofluor white and refed onto a YPD microscope pad. (A) Percentage of mother or daughter cells with vesicular mitochondria …

https://doi.org/10.7554/eLife.35685.014
Figure 4—source data 1

Cell category scoring for each replicated experiment in Figure 4 panel A, B to D, E and F.

https://doi.org/10.7554/eLife.35685.015
Figure 5 with 1 supplement
Actin, proteasome and mitochondrial network reorganization.

WT cells expressing Ilv3-RFP and Abp1-3xGFP (actin) or Scl1-GFP (proteasome) were grown 7 d in YPD and refed on a YPD microscope pad. (A) Histograms show the percentage of individual cells …

https://doi.org/10.7554/eLife.35685.016
Figure 5—source data 1

Cell category scoring for each replicated experiment in Figure 5 panel A, B and D.

https://doi.org/10.7554/eLife.35685.019
Figure 5—figure supplement 1
Actin and Proteasome localization in WT and rho zero cells, cell volume variation according to mitochondrial network organization.

(A) Actin phalloidin staining of WT prototroph rho zero cells after 24 hr of culture in YPD. The image is a superposition of the DIC and the RFP channel. (B) Proteasome (Scl1-GFP) localization in WT …

https://doi.org/10.7554/eLife.35685.017
Figure 5—figure supplement 1—source data 1

Cell volume measurements in daughter and mother cells depending on their mitochondrial network organization (panel C).

https://doi.org/10.7554/eLife.35685.018

Tables

Key resources table
ResourceDesignationSource or referenceIdentifiers
SoftwareImageJDepartment of Health and Human Services, NIDARRID:SCR_003070
SoftwareGraphPad Prism 5GraphPad Software, Inc. La Jolla, USARRID:SCR_015807

Additional files

Supplementary file 1

Table with the genotype of the strains used in this study.

https://doi.org/10.7554/eLife.35685.020
Transparent reporting form
https://doi.org/10.7554/eLife.35685.021

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