Figure 8.

(A) Robust FANCI ubiquitination requires its DNA binding activity but not FANCL. Reactions with FANCI or one of its mutants (0.5 or 1 μM), and with or without DNA (45-nM ϕX virion), were incubated for 18 h (fluorescent-stained gel) or 3 h (western blot). The asterisk indicates a degradation band of FANCI. (B) The FANCI R1285X and R1285Q mutants are ubiquitinated well. The experiments were performed under the same conditions as in panel (A). (C) Ubiquitination reactions were performed with and without UBE2T or the C86A catalytically defective mutant. Analysis was as described for panel (A). (D) Ubiquitination reactions were performed with or without ϕX replicative form DNA (34 nM), FANCL or the W212A/L214A (WL/AA) mutant variant, as indicated. Analysis was as described for panel (A).