Extended Data Fig. 6: Acetyl-CoA Synthase (ACSS2) generates acetyl-CoA for H3K18ac.
From: Nuclear position and local acetyl-CoA production regulate chromatin state
(a) Cross-sections of wing discs stained with TMRM show high mitochondrial membrane potential in the proximal folds, indicating a broader pattern than high H3K18ac. Discs incubated for 30 min in explant cultures with TMRM. (n = 13 discs). (b) FABO produces acetyl-CoA inside mitochondria which is converted to acetate or citrate for export from this organelle. Acetate and citrate, in turn, can be used to generate nuclear acetyl-CoA by acetyl-CoA synthase (AcCoAS/ACSS2) or ATP citrate lyase (ATPCL/ACLY). Nuclear acetyl-CoA is used by acetyltransferases such as nejire (nej) to acetylate histones. (c-d) ACLY is not required for H3K18ac. ACLY knockdown, validated by ACLY immunostaining (c), does not impact H3K18ac levels (d). Temperature-sensitive knockdown of ACLY was induced for 2 days at 29 °C in the posterior compartment (IFP+). (c, n = 9 discs; d, n = 11 discs). (e-f) Acetyl-CoA required for acetylation of H3K18 is not derived from citrate, as citrate treatment (10 mM) does not rescue loss of H3K18ac upon inhibition of FABO inhibition (500 μM etomoxir). Discs incubated with the indicated compounds for 2 h in explant cultures. Representative images in (e), quantified in (f). Statistical significance by Kruskal-Wallis test with Dunn’s multiple comparisons test. (g-h) ACSS2 provides acetyl-CoA for H3K18ac. Inhibition of ACSS2 (20 μM ACSS2 inhibitor, ACSS2i) for 2 h in explant cultures causes loss of H3K18ac. Representative images in (g), quantified in (h). Statistical significance by Mann-Whitney test (two-sided). (i-j) ACSS2 is required for acetate (10 mM) to rescue the etomoxir-induced (500 μM) loss of H3K18ac. Discs incubated for 2 h in explant cultures with indicated compounds ± ACSS2i (20 μM). Representative images in (i), quantified in (j). Statistical significance by Kruskal-Wallis test with Dunn’s multiple comparisons test. Box plots: center line (median), box limits (1st and 3rd quartiles), whiskers (outer data points).