Fig. 2: N-terminal domain of p14ARF is essential for SUMO conjugation.
From: SUMOylation of the lysine-less tumor suppressor p14ARF counters ubiquitylation-dependent degradation
A In vitro SUMOylation of 35S-methionine-labeled p14ARF protein after incubation or not with amino group blocking agent sulfo-NHS-acetate. B Modification of transfected p14ARF-GFP protein by SUMO2 in HEK-293 cells co-transfected with His6-SUMO2, UBC9 and pcDNA or His6-SUMO2, UBC9 and Flag-hNaa60. Arrow indicates unmodified p14ARF protein and arrowhead indicates SUMOylated p14ARF. C Scheme of constructs encoding C-terminal GFP-tagged deletion mutants of p14ARF. D Modification of p14ARF(1–28)-GFP or p14ARF(21-132)-GFP protein by SUMO2 in HEK-293 cells transfected with His6-SUMO2 and UBC9. E Modification of p14ARF(31-132)-GFP or p14ARF(36-132)-GFP protein by SUMO2 in HEK-293 cells transfected with His6-SUMO2 and UBC9. WCE whole cell extracts.
