Figure 5: SRSF3 is neddylated at Lys85 under arsenite stress.

(a) Schematic representation of SRSF3 protein domains and positions of lysine residue. SRSF3 contains 2 domains, one RRM and one RS domain and has 5 lysine residues positioned at 11, 23, 85, 146 and 164 (highlighted by underline). (b) SRSF3 is neddylated at Lys85 under arsenite stress. His-NEDD8 was co-transfected with empty vector or Flag–SRSF3 WT and series of single lysine mutant (11, 23, 85, 146, 164), double lysine mutant (11/23, 23/85, 146/164) or null mutant (all lysine mutant) as indicated and treated with 0.5 mM arsenite for 30 min. Cells were then lysed under denaturing condition, affinity-purified with Ni-NTA agarose beads and the precipitates were subjected to western analysis using anti-Flag antibody.