Fig. 3: Switching occurs between protein synthesis (HOC) and autophagy (LOC). | Nature Communications

Fig. 3: Switching occurs between protein synthesis (HOC) and autophagy (LOC).

From: Eukaryotic cell biology is temporally coordinated to support the energetic demands of protein homeostasis

Fig. 3

a Model: cells accumulate carbohydrates (CH2O)n, amino acids and osmolytes during LOC and consume/export them in HOC to sustain translational bursts and maintain osmostasis. Replete stores increase H+ export, a pH-dependent checkpoint activating TORC1 and releasing BMC proteins. HOC ends when stores are exhausted, see Fig. 5e and Supplementary Table 1 for more details. b Puromycin incorporation assay and immunoblot for TORC1 activation (phospho-Rps6, Ser235/236) reveal translational bursting in HOC (n = 3 biologically independent samples, TWAINT: two-way ANOVAINTERACTION, total protein loading control). c Immunoblots for cleaved/full-length Pgk1-GFP reveal increased autophagy during LOC (n = 3 biologically independent samples, OWA one-way ANOVA, total protein loading control). d Differential variation in vacuole volume vs. surface area:volume ratio (two-sided unpaired t-test, n = 7 independent experiments for HOC and LOC, n > 68 cells per image). Data throughout presented as mean ± SEM where *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. e Acute inhibition of protein synthesis (CHX, 25 µg mL−1 cycloheximide or TORC1 activity 200 nM rapamycin) during HOC immediately terminates HOC and abolishes the YRO, representative OCR and H+-export traces are shown. Source data are provided as a Source Data file.

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