Figure 4: Formation of the (MHF1–MHF2)2 tetramer is essential for MHF function in vivo.
From: The structure of the FANCM–MHF complex reveals physical features for functional assembly
(a) Dimeric complex of MHF1 is disrupted by His71A and Asp81A mutations. Immunoprecipitation results show that wild-type Flag-MHF1 pulls down GFP–MHF1 (lane 3) but not MHF1H71A/D81A mutant (lane 4). (b) (MHF1–MHF2)2 tetramer is essential for a stable localization to centromere. GFP–MHF1 is readily apparent as it co-localizes with centromere mark ACA (anti-centromere autoantibody; upper panel merge). GFP–MHF1H71A/D81A failed to localize to the centromere as the GFP signal was not concentrated to the centromere (lower panel). (c) Integral (MHF1–MHF2)2 is essential for a stable localization to centromere. GFP–MHF2 is readily apparent as it colocalizes with centromere mark ACA (upper panel merge). GFP–MHF2D80A/F81A failed to localize to the centromere as the GFP signal was not concentrated to the centromere (lower panel). (d) MHF1–MHF2 heterodimer is perturbed by mutation of MHF2D80A/F81A. Western blot analysis results show that MHF1 pulls down GFP–MHF2 (lane 3) but not MHF2D80A/F81A mutant (lane 4).
