(A) ARIH1 domains.
(B) Sites of Ala mutations as spheres on structure of autoinhibited ARIH1 (Duda et al., 2013), colored by effect on *UB transfer from UBCH7 via ARIH1/neddylated CRL1FBW7ΔD to pCyE (in Figure S4): gray, normal; green, hyperactive; blue, ligation defect/accumulation of normally transient thioester-bonded ARIH1~UB intermediate; raspberry, marginal defect; red, strong defect.
(C) Most defective (red) and ligation-impaired (blue) ARIH1 Ala scan mutants in pulse-chase assays monitoring entire pathway: *UB transfer from UBCH7 via the indicated ARIH1 mutants and neddylated CRL1FBW7ΔD to pCyE. Ligation-impaired mutants are scored by appearance and dissipation of ARIH1~UB thioester intermediate.