Fig. 4: HAG704 HAT activity on H4K5 depends on ACL.
From: ACL and HAT1 form a nuclear module to acetylate histone H4K5 and promote cell proliferation
a Analysis of histone acetylation levels in hag704-1 and hag704-2 mutants compared with the wild type (WT-1 and WT-2) plants by immunoblotting. Histone proteins extracted from 6 DAF seeds were tested. The histone acetylation antibodies used in the tests are indicated on the left. Total histone extracts were used for the analysis. Two replicates are shown. The immunoblot signals were quantified using ImageJ. Relative signals (to the control WT1 signal set at 1) are indicated below the bands. b In vitro HAG704 histone acetyltransferase activity assays. His-tagged histone H3 or H4 proteins were produced alone or together with and HAG704 protein in co-transfected tobacco leaf cells. After purification with His resins, the acetylation levels of the His-tagged histone H3 or H4 proteins were analyzed by immunoblotting with the antibodies indicated on the left. Anti-H3 and anti-H4 antibodies were used as loading controls. c ACL stimulated the HAG704 histone acetyltransferase activity at H4K5. His-tagged H4 protein was produced together with or without combinations of HAG704, ACLA2 and ACLB-FLAG in tobacco leaf cells. The acetylation level of purified histone H4 was analyzed by immunoblots using the H4K5ac and H4K16ac antibodies. ACLA2, HAG704, and ACLB were detected by anti-ACLA2, anti-HAG704, and anti-FLAG antibodies respectively. Anti H4 antibody was used as loading controls. Two replicates are shown. Immunoblotting results were quantified using ImageJ. Relative signals (to controls, i.e., in the absence of HAG704, set at 1) are indicated below the bands. Source data are provided as a Source Data file.
