FIGURE 3.

Deacetylase activity is not required for HDAC2-induced mRNA translation of reporter genes. A, overexpression of wild-type HDAC2 and HDAC2-H142A mutant induces cap-dependent translation of luciferase reporter gene. The HCT-116 cells were transfected with empty vector (VC), wild-type (WT) HDAC2, or HDAC2-H142A mutant along with luciferase cDNA reporter. 48 h after transfection, the transfected cells were harvested for luciferase assay. The absolute luciferase relative light units values are in the 10⁷–10⁸ range. Statistics were performed by one-way analysis of variance followed by Tukey's multiple comparison test using the data obtained from three independent experiments. B, butyrate does not inhibit basal translation rate. The HCT-116 cells were transfected with luciferase cDNA reporter, and 24 h after transfection, the transfected cells were treated with butyrate at the indicated concentration for 24 h. C, butyrate treatment does not suppress HDAC2-medited induction of luciferase activity. Statistics were performed by one-way analysis of variance followed by Tukey's multiple comparison test using the data obtained from three independent experiments. Error bars in A–C indicate S.E. D, overexpression of HDAC2 on cap-dependent translation of c-Myc-tagged EYFP and IRES-dependent translation of HA-tagged ECFP. The HCT-116 cells were transfected with HDAC2 or empty vector along with bicistronic vector. Cap-dependent translation of c-Myc-EYFP protein was evaluated by immunoblotting with anti-c-Myc. IRES-dependent translation of HA-ECFP was evaluated by immunoblotting with anti-HA. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.