Fig. 2: PKD1 phosphorylates KAT7 at Thr97/Thr331 in vitro and in vivo. | Cell Death Discovery

Fig. 2: PKD1 phosphorylates KAT7 at Thr97/Thr331 in vitro and in vivo.

From: Protein kinase D1 phosphorylation of KAT7 enhances its protein stability and promotes replication licensing and cell proliferation

Fig. 2

a HEK293T cells were co-transfected Flag-KAT7 with or without HA-PKD1-CA. An IP assay was carried out using anti-Flag antibody followed by immunoblotting with anti-pMOTIF antibody, anti-phosphothreonine, anti-phosphoserine or anti-Flag antibodies, respectively. b HeLa cells were transfected with or without HA-PKD1-CA. An IP assay was carried out using anti-KAT7 antibody and immunoblotted with the indicted antibodies. c Bacterially expressed and purified GST (negative control) or GST-KAT7 was incubated with purified HA-PKD1-CA in a kinase reaction. The reaction products were separated by SDS-PAGE and immunoblotted with the anti-GST and the other indicated antibodies. d HEK293T cells were co-transfected Flag-KAT7 with HA-PKD1-CA or HA-PKD1-KD. An IP assay was performed and immunoblotted with the indicted antibodies. e Bacterially expressed and purified GST-KAT7 was incubated with purified HA-PKD1-CA or HA-PKD1-KD in a kinase reaction. The reaction products were separated by SDS-PAGE and immunoblotted with the indicated antibodies. f HEK293T cells were co-transfected with Flag-KAT7 and HA-PKD1-CA and treated with or without Gö6976. An IP assay and subsequent immunoblotting were performed as above. g HEK293T cells were cotransfected with WT-KAT7 or its mutants KAT7-T97A, T331A, or T97/331A and HA-PKD1-CA. An IP assay was performed and immunoblotted with the indicated antibodies. h Purified GST-tagged WT-KAT7 or its mutants were incubated with or without HA-PKD1-CA in a kinase reaction, and subsequent steps were performed as above.

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