Fig. 3: HADC7 is the deacetylase for ALKBH5 acetylation at K235. | Nature Communications

Fig. 3: HADC7 is the deacetylase for ALKBH5 acetylation at K235.

From: K235 acetylation couples with PSPC1 to regulate the m6A demethylation activity of ALKBH5 and tumorigenesis

Fig. 3

a HADC7 overexpression reduced ALKBH5 acetylation at K235. HeLa cells were transfected with the indicated plasmids together with the ALKBH5-HA vector; ALKBH5-HA was IPed using anti-HA antibody, and K235 acetylation in the IPed ALKBH5-HA was determined using the anti-pan acetylated lysine antibody. b, c ALKBH5 interacted with HDAC7. The ALKBH5-HA and HDAC7-FLAG plasmids were cotransfected into HeLa cells, and the HDAC7-FLAG (b) and ALKBH5-HA (c) complexes were co-IPed using anti-FLAG and anti-HA antibodies, respectively. ALKBH5-HA and HDAC7-FLAG were detected. d Silencing of HDAC7 increased the endogenous K235 acetylation of ALKBH5. HeLa cells were transfected with anti-HDAC7 siRNAs, and K235 acetylation was determined. e HDAC7 overexpression decreased the endogenous K235 acetylation of ALKBH5. HeLa cells were transfected with the indicated dose of HDAC7 plasmid, and K235 acetylation was determined. f HDAC7 directly deacetylated ALKBH5 at K235 in a dose-dependent manner in the in vitro deacetylation reaction. Immunopurified wild-type ALKBH5 containing K235-acetylated ALKBH5 stimulated by KAT8 overexpression was incubated with the indicated dose of immunopurified HADC7, and ALKBH5 acetylation at K235 was determined using anti-Ac K235 and anti-pan acetylated lysine antibodies. Source data are provided as a Source Data file.

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