Figure 2
From: RUNX family members are covalently modified and regulated by PIAS1-mediated sumoylation
RUNX3 is sumoylated on lysine 148 that is evolutionarily conserved. (a) Schematic representation of human RUNX3. Lysine residues around the Runt domain are indicated. To identify the region required for sumoylation, Myc-RUNX3 deletion mutants were coexpressed with HA-PIAS1 and FLAG-SUMO1 in HEK293 cells. RUNX3 sumoylation was analyzed by IB. (b) Each lysine (K) residue of RUNX3 was individually mutated to arginine (R), and each Myc-RUNX3-KR mutant was coexpressed with HA-PIAS1 and FLAG-SUMO1 in HEK293 cells. RUNX3 sumoylation was analyzed by IB. (c) Amino acid sequence around the RUNX3 sumoylation site (K148) was compared among RUNX3 family members. c, chicken; d, Drosophila melanogaster; h, human; m, mouse; r, rat; x, Xenopus laevis; z, zebra fish. The conserved lysine residues used for RUNX3 sumoylation are indicated by shadow. GenBank accession numbers for each protein: human RUNX1, NP_001001890.1; human RUNX2, NP_001265407; human RUNX3, NP_004341.1; mouse, NP_062706.2; rat, NP_569109.1; chicken, XP_001232978, Xenopus: NP_001182313.1; zebrafish, NP_571679.2; Drosophila, AAC47196. (d) Myc-RUNX1-WT and Myc-RUNX1-K144R were coexpressed with HA-PIAS1 and FLAG-SUMO1 in HEK293 cells. RUNX1 sumoylation was analyzed by IB. (e) Myc-RUNX2-WT and Myc-RUNX2-K187R were coexpressed with HA-PIAS1 and FLAG-SUMO1 in HEK293 cells. RUNX2 sumoylation was analyzed by IB. (f) Myc-lz-WT and Myc-lz-372 were coexpressed with HA-dPias and FLAG-dSumo in HEK293 cells. The lz sumoylation was analyzed by IB.
