Open Access Research article

Uncovering by Atomic Force Microscopy of an original circular structure at the yeast cell surface in response to heat shock

Flavien Pillet12, Stéphane Lemonier123, Marion Schiavone12456, Cécile Formosa1278, Hélène Martin-Yken456, Jean Marie Francois456* and Etienne Dague123*

Author Affiliations

1 CNRS, LAAS, 7 avenue du colonel Roche, F-31077 Toulouse, France

2 Université de Toulouse, UPS, INSA, INP, ISAE, LAAS, F-31077 Toulouse, France

3 CNRS, ITAV-USR 3505, F31106 Toulouse, France

4 Université de Toulouse, INSA, UPS, INP, 135 avenue de Rangueil, F-31077 Toulouse, France

5 INRA, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31077 Toulouse, France

6 CNRS, UMR5504, F-31400 Toulouse, France

7 CNRS, UMR 7565, SRSMC, Vandoeuvre-lès-Nancy, France

8 Université de Lorraine, UMR 7565, Faculté de Pharmacie, Nancy, France

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BMC Biology 2014, 12:6  doi:10.1186/1741-7007-12-6

Published: 27 January 2014

Additional files

Additional file 1: Figure S1:

The accumulation of trehalose is correlated with survival of cells under heat stress condition. Comparison of trehalose accumulation in the wild-type yeast BY4741 and the defective mutants wsc1Δ and bck1Δ. Control (full bar) and heat-shocked condition (hachured bar) are represented.

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Additional file 2: Table S1:

Evaluation of viability by blue methylene test. The percentage of mortality was evaluated before and after heat shock with the defective mutants wsc1 and bck1, and the wild-type yeast with or without nitrogen starvation during 72 h.

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Additional file 3: Figure S2:

Young modulus increase with heat-shock. Distribution of Young modulus values calculate with 19 elasticity maps (ncurves = 19443) from individual yeasts unstressed (A), in comparison with 15 elasticity maps (ncurves = 15307) from individual yeasts heat-shocked at 42°C (B). YM medians were indicated on diagrams and calculated from fits in gauss model (red curves). (C) Statistic unpaired t test between averages and standard deviations calculated from young modulus values. The 3 asterisks shown significant differences between elasticity of unstressed yeasts (full bar) and heat-shock yeasts (hachured bar) at the P value < 0.0001.

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Additional file 4: Figure S3:

The absence of F-Actin prevent the formation CS. AFM high resolution images of wild-type cells after heat shock in absence (A) or in presence of 200 μM Latrunculin A (B). Cells incubated 1 hr at 30°C with 200 μM of Latrunculin A (C).

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Additional file 5: Figure S4:

The formation of CS require budding process. High-resolution deflection images of wild-type incubate 72 h at 30°C in nitrogen starvation, without (A) or with heat shock 1 hr at 42°C (B).

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Additional file 6: Figure S5:

The heat-induced formation of the cell surface circular structure is abolished in mutants defective in the budding process. High-resolution AFM deflection images of bni1Δ(A), chs3Δ(B) and chs1Δ(C) mutants after heat shock.

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Additional file 7: Figure S6:

The CWI controls the stiffness of the cell wall and the formation of the cell surface circular structure in response to heat shock. High-resolution AFM deflection images of wild-type cell (A), wsc1Δ(B) and bck1Δ(C) cell defective in the CWI pathway imaged after 1 hr of incubation at 42°C.

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Additional file 8: Figure S7:

The stiffness of wsc1Δ unstressed was similar to wild-type yeast exposed at 42°C during 1 h. Distribution of Young modulus values calculate with 4 elasticity maps (n = 4096) from individual wsc1Δ yeasts unstressed.

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Additional file 9: Table S2:

Chitin rate was similar in wsc1Δ mutant with or without heat-shock at 42°C. Carbohydrate composition of wsc1Δ mutant was determinated by acid hydrolysis and enzymatic method and expressed in μg/mg of cell wall dry mass.

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Additional file 10: Figure S8:

Yeast immobilization on PDMS stamp. (A) AFM height image of a PDMS stamp containing some immobilized yeasts. The z range is 2.5 μm. (B) 3D projection associated to the height image.

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